Compositions for drug administration

ABSTRACT

The present invention provides compositions and methods and for increasing the bioavailability of therapeutic agents in a subject. The compositions include at least one alkyl glycoside and at least one therapeutic agent, wherein the alkylglycoside has an alkyl chain length from about 10 to about 16 carbon atoms.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 15/890,131 filed Feb. 6, 2018, now pending. The disclosure of theprior application is considered part of and is incorporated by referencein the disclosure of this application.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates to generally to compositions containing apharmaceutically active ingredient and an alkylsaccharide.

Background Information

Therapeutic agents are often combined with various surfactants. Yet,surfactants are frequently irritating to the skin and other tissues,including mucosal membranes such as those found in the nose, mouth, eye,vagina, rectum, esophagus, intestinal tract, and the like. Manysurfactants also cause proteins to denature, thus destroying theirbiological activity. Another serious limitation to the development anduse of such agents is the ability to deliver them safely,non-invasively, efficiently and stably to the site of action. Therefore,an ideal enhancing surfactant will stabilize the therapeutic agent, benon-toxic and non-irritable to the skin or mucosal surfaces, haveantibacterial activity, and enhance the passage or absorption of thetherapeutic agent through various membrane barriers without damaging thestructural integrity and biological function of the membrane andincrease bioavailability of the agent.

In spite of the many attractive aspects of peptides and proteins aspotential therapeutic agents, their susceptibility to denaturation,hydrolysis, and poor absorption in the gastrointestinal tract makes themunsuitable for oral administration, typically requiring administrationby injection. This remains a major shortcoming. Compared to smallmolecule drugs, peptides are considerably less stable. Careful attentionmust be paid to formulation and storage to avoid unwanted degradation.Some proteins, particularly proteins with substantially non-naturallyoccurring amino acid sequences can be immunogenic. Upon injection,immune cells may be recruited to the site of injection and a humoral orcellular immune response may be induced. Aggregated peptides are knownto be more prone to eliciting an immunogenic response than monomers.This may be avoided to a greater to or lesser extent if the peptide canbe directly absorbed from the gastrointestinal tract into systemiccirculation. Therefore, while the range of clinical indications fortherapeutic proteins and peptides is quite broad, the actual number ofsuch therapeutics in general use today is quite small compared to thenumber of chemically synthesized and orally active pharmaceuticalscurrently on the market. In recent years, development of a large classof alkylsaccharide delivery enhancement agents, for example, moleculesthat provide intranasal bioavailabilities, comparable to those achievedby injection have been investigated. While recent developments inintranasal delivery for proteins and peptides are creating new andexpanded opportunities for practical clinical uses of peptides,proteins, and other macromolecular therapeutics, few, if any, peptidesappear to be administrable orally due to unacceptably low oralbioavailability. A number of studies have been conducted to demonstrateoral bioavailability for a variety of peptide drugs. These studies useda variety of absorption enhancers as well as physical processes such asmicronization. For example among formulations specifically optimized fororal delivery, insulin exhibited only 3% oral bioavailability (Badwin etal., 2009). Calcitonin exhibited only 0.5-1.4% oral bioavailability(Bucklin 2002). Parathyroid hormone has been shown to exhibit 2.1% oralbioavailability (Leone-Bay et al., 2001). There are two principalbiochemical problems limiting the oral absorption of peptides. The firstrelates to the susceptibility of peptides to hydrolysis in thegastrointestinal tract. The second relates to intrinsically poorabsorption across the intestinal mucosal membrane.

Incorporation of non-standard amino acids into peptide sequences hasbeen shown to reduce hydrolysis or slow metabolism for some peptides.Non-standard aminoacyl residues have been incorporated into a number ofdrugs for this purpose allowing the drugs to remain active for a longerperiod of time than otherwise possible. Non-standard amino acids arethose amino acids that are not among the 22 naturally occurring L-aminoacids found in proteins. There exist a vast number of non-standard aminoacids that may be considered for such use in either the D or Lconfiguration. A few examples include, but are not limited to,allylglycine, (2S,3R,4S)-α-(carboxycyclopropyl)glycine,α-cyclohexylglycine, C-propargylglycine, α-neopentylglycine,α-cyclopropylglycine, N-lauroylsarcosine sodium salt,N-(4-hydroxyphenyl)glycine, N-(2-furoyl)glycine, naphthylglycine,phenylglycine, lanthionine, 2-aminoisobutyric acid, dehydroalanine,gamma-aminobutyric acid.

Some specific examples of non standard amino acids used in drugs includeD-4-hydroxyphenylglycine which is incorporated into the antibacterialdrug Amoxicillin, D-phenylglycine which is incorporated into theantihypertensive drug Enalapril, and (2R,3S)-phenylisoserine which isincorporated into the antineoplastic drug Taxol.

In the case of peptide drugs, D-2-Naphthylalanine is incorporated intothe endometriosis drug Nafarelin. The D-isomers of naturally occurringL-amino acids are frequently used to increase stability of peptidedrugs. Examples of D-amino acid stabilized peptides include theanti-obesity peptide D-Leu-OB3 (Lee et al., 2010) and the CCR5 anti-HIVdrug D-ala-peptide T (DAPTA) (Ruff et al., 2001) among others.

Enzymatic hydrolysis in the gastrointestinal tract may also be reducedor eliminated by addition of specific enzyme inhibitors such asbacitracin, bestatin, amastatin, boroleucin, borovaline, aprotinin, andtrypsin inhibitor among others.

Alkylsaccharides have been demonstrated to enhance oral absorption ofsmall molecules, and peptides, when presented as aqueous solutions ofthe alkylsaccharide and the small molecule or peptide. In suchsolutions, the concentration of the alkylsaccharide is higher than thecritical micelle concentration (CMC). An example includes the oraldelivery of octreotide in aqueous solution usingdodecyl-beta-D-maltoside as the alkylsaccharide absorption enhancer.Another example is oral delivery of a leptin-related synthetic peptideinsulin sensitizer using an aqueous dodecyl maltoside solution. Yetanother example is oral delivery of exenatide and pramlintide usingdodecyl-beta-D-maltoside in an aqueous solution as the alkylsaccharideabsorption enhancer. Yet another example is oral delivery of heparinusing tetradecyl maltoside in an aqueous solution as the alkylsaccharide absorption enhancer (see for example, Maggio and Grasso,Regulatory Peptides 167 (2011) 233-238; Novakovic, et al., Peptides, 43(2013) 167-163; Leinung M C et al., Regulatory Peptides 179:33-38(2012); Yang, et al., (2005) Journal of Drug Targeting, 13:1, 29-38).

SUMMARY OF THE INVENTION

The present invention is based, in part, on the development of atherapeutic composition containing a drug enhancing agent useful forincreasing the absorption and bioavailability of the drug, while at thesame time avoiding various adverse toxic effects of drug. In particular,the drug enhancing agents of the invention contain a non-toxicsurfactant consisting of at least an alkyl glycoside and/or saccharidealkyl ester. One advantage of the therapeutic compositions of theinvention is that they permit administration and delivery of thetherapeutic agents with high bioavailabilities at concentrations ofenhancing agents that are dramatically below their so-called “noobservable adverse effect levels” (their NOAEL's). Accordingly, thepresent invention provides compositions, including alkyl glycosidesand/or saccharide alkyl esters and a therapeutic agent (e.g., smallmolecule organic drug molecules, low molecular weight peptides such asExenatide, GLP-1 and the like, proteins, and non-peptide therapeuticpolymers such as low molecular weight heparin and inhibitory RNA),methods of administering and using the compositions e.g., via the oral,ocular, nasal, nasolacrimal, inhalation or pulmonary, oral cavity(sublingual or Buccal cell) or cerebral spinal fluid (CSF) deliveryroute, and methods of ameliorating a disease state in a subject byadministration of such compositions.

Previously, no examples of the use of alkylsaccharides as absorptionenhancers in solid dosage forms such as tablets has been demonstrated.While alkylsaccharides such as dodecyl maltoside, n-tetradecyl maltosidehave relatively high levels of solubility in aqueous solution, theydissolve only very slowly and because a significant portion of themolecules are comprised of the hydrophobic linear alkyl chains. In orderto ensure complete dissolution of these alkylsaccharides in aqueoussolution, the aqueous solution is mildly heated and the containerholding the aqueous media and the alkylsaccharide is either stirred oragitated continuously for up to 15 to 30 min. in order to ensurecomplete dissolution of the alkylsaccharide. As a result, it was notexpected that solid dosage forms containing an alkylsaccharide and thedrug substance would benefit from the absorption enhancing effect of thealkylsaccharide since the slow dissolution of the alkylsaccharide in theaqueous gastrointestinal contents would not be expected to produce asufficiently high concentration or comparable concentration to thatachieved in the aqueous formulations before the drug substance and thealkylsaccharide molecules diffuse away from each other throughout thegastrointestinal contents diluting the relative concentrations of each.Surprisingly, solid dosage forms comprising an alkylsaccharide, and apharmacologically active substance, along with other inactive excipientsformed into a tablet were found to provide a substantial increase inoral bioavailability of the pharmacologically active substance, as shownin the examples. In addition to active pharmaceutical substance andalkylsaccharide absorption enhancer, other inactive excipients mayinclude by way of example candelilla wax, hypromellose, magnesiumstearate, microcrystalline cellulose, polyethylene glycol, povidone, andtitanium dioxide.

In one aspect, the present invention relates to a surfactant compositionhaving at least one alkyl glycoside and/or at least one saccharide alkylester, and when admixed, mixed or blended with a therapeutic agent, adrug, or biologically active compound, the surfactant stabilizes thebiological activity and increases the bioavailability of the drug.

Accordingly, in one aspect, the invention provides a therapeuticcomposition having at least one biologically active compound and atleast one surfactant, wherein the surfactant further consists of atleast one alkyl glycoside and/or saccharide alkyl ester or sucrose esterand wherein the therapeutic composition stabilizes the biologicallyactive compound for at least about 6 months, or more, and from about 4°C. to about 25° C.

The invention also provides a method of administering a therapeuticcomposition having a surfactant including at least one alkyl glycosideand/or saccharide alkyl ester admixed, mixed, or blended with at leastone therapeutic agent, or a drug, or biologically active compound, andadministered or delivered to a subject, wherein the alkyl has from about10 to 24, 10 to 20, 10 to 16, or 10 to 14 carbon atoms, wherein thesurfactant increases the stability and bioavailability of thetherapeutic agent.

In yet another aspect, the invention provides a method of increasingabsorption of a low molecular weight compound into the circulatorysystem of a subject by administering the compound via the oral, ocular,nasal, nasolacrimal, inhalation or pulmonary, oral cavity (sublingual orBuccal cell), or CSF delivery route when admixed, mixed or blended withan absorption increasing amount of a suitable surfactant, wherein thesurfactant is a nontoxic and nonionic hydrophobic alkyl joined by alinkage to a hydrophilic saccharide. Such low molecular weight compoundsinclude but are not limited to, nicotine, interferon, PYY, GLP-1,synthetic exendin-4, parathyroid hormone, human growth hormone, or asmall organic molecule. Additional low molecular weight compoundsinclude antisense oligonucleotides or interfering RNA molecules (e.g.,siRNA or RNAi).

The present invention also provides a method of treating diabetesincluding administering to a subject in need thereof via the oral,ocular, nasal, nasolacrimal, inhalation or pulmonary, or oral cavity(sublingual or Buccal cell), a blood glucose reducing amount of atherapeutic composition, for example, an incretin mimetic agent or afunctional equivalent thereof, and an absorption increasing amount of asuitable nontoxic, nonionic alkyl glycoside having a hydrophobic alkylgroup joined by a linkage to a hydrophilic saccharide, therebyincreasing the absorption of incretin mimetic agent or insulin andlowering the level of blood glucose and treating diabetes in thesubject.

The present invention also provides a method of treating congestiveheart failure in a subject including administering to the subject inneed thereof via the oral, ocular, nasal, nasolacrimal, or inhalationdelivery route, a therapeutically effective amount of a compositioncomprising a GLP-1 peptide or a functional equivalent thereof, and anabsorption increasing amount of a suitable nontoxic, nonionic alkylglycoside having a hydrophobic alkyl joined by a linkage to ahydrophilic saccharide, thereby treating the subject.

In another aspect, the invention provides a method of treating obesityor diabetes associated with obesity in a subject comprisingadministering to a subject in need thereof via the oral, ocular, nasal,nasolacrimal, inhalation or CSF delivery route, a therapeuticallyeffective amount of a composition comprising a PYY peptide or afunctional equivalent thereof, and an absorption increasing amount of asuitable nontoxic, nonionic alkyl glycoside having a hydrophobic alkyljoined by a linkage to a hydrophilic saccharide, thereby treating thesubject.

In another aspect, the invention provides a method of increasingabsorption of a low molecular weight therapeutic compound into thecirculatory system of a subject by administering via the oral, ocular,nasal, nasolacrimal, inhalation or CSF delivery route the compound andan absorption increasing amount of a suitable nontoxic, nonionic alkylglycoside having a hydrophobic alkyl group joined by a linkage to ahydrophilic saccharide, wherein the compound is from about 1-30 kD, withthe proviso that the compound is not insulin, calcitonin, or glucagonwhen the route of administration is oral, ocular, nasal, ornasolacrimal.

The present invention also provides a method of increasing absorption ofa low molecular weight therapeutic compound into the circulatory systemof a subject by administering via the oral, ocular, nasal, nasolacrimal,inhalation or pulmonary, oral cavity (sublingual or Buccal cell) or CSFdelivery route the compound and an absorption increasing amount of asuitable nontoxic, nonionic alkyl glycoside having a hydrophobic alkyljoined by a linkage to a hydrophilic saccharide, wherein the compound isfrom about 1-30 kilo Daltons (kD), with the proviso that the subjectdoes not have diabetes when delivery is via the oral, ocular, nasal ornasolacrimal routes.

In one aspect of the invention, there is provided a pharmaceuticalcomposition having a suitable nontoxic, nonionic alkyl glycoside havinga hydrophobic alkyl group joined by a linkage to a hydrophilicsaccharide in combination with a therapeutically effective amount ofExenatide (exendin-4) in a pharmaceutically acceptable carrier.

In one aspect, the invention provides a pharmaceutical compositionhaving a suitable nontoxic, nonionic alkyl glycoside having ahydrophobic alkyl group joined by a linkage to a hydrophilic saccharidein combination with a therapeutically effective amount of GLP-1 in apharmaceutically acceptable carrier.

In one aspect, the invention provides a pharmaceutical compositionhaving a suitable nontoxic, nonionic alkyl glycoside having ahydrophobic alkyl group joined by a linkage to a hydrophilic saccharidein combination with a therapeutically effective amount of nicotine in apharmaceutically acceptable carrier.

In one aspect, the invention provides a pharmaceutical compositioncomprising a suitable nontoxic, nonionic alkyl glycoside having ahydrophobic alkyl group joined by a linkage to a hydrophilic saccharidein combination with a therapeutically effective amount of interferon ina pharmaceutically acceptable carrier.

In one aspect, the invention provides pharmaceutical composition havinga suitable nontoxic, nonionic alkyl glycoside having a hydrophobic alkylgroup joined by a linkage to a hydrophilic saccharide in combinationwith a therapeutically effective amount of PYY in a pharmaceuticallyacceptable carrier.

In one aspect, the invention provides a pharmaceutical compositionhaving a suitable nontoxic, nonionic alkyl glycoside having ahydrophobic alkyl group joined by a linkage to a hydrophilic saccharidein combination with a therapeutically effective amount of parathyroidhormone in a pharmaceutically acceptable carrier.

In one aspect, the invention provides a pharmaceutical compositionhaving a suitable nontoxic, nonionic alkyl glycoside having ahydrophobic alkyl group joined by a linkage to a hydrophilic saccharidein combination with a therapeutically effective amount of a peptidehaving a molecular weight of about 1-75 kD in a pharmaceuticallyacceptable carrier, with the proviso that the peptide is not insulin,calcitonin, and glucagon.

In one aspect, the invention provides a pharmaceutical compositionhaving a suitable nontoxic, nonionic alkyl glycoside having ahydrophobic alkyl group joined by a linkage to a hydrophilic saccharidein combination with a therapeutically effective amount erythropoietin ina pharmaceutically acceptable carrier.

In one aspect, the invention provides a pharmaceutical compositionhaving a therapeutically effective amount of an oligonucleotide incombination with an absorption increasing amount of an alkylglycoside.The oligonucleotide can be an antisense oligonucleotide or interferingRNA molecules, such as siRNA or RNAi. The oligonucleotide typically hasa molecular weight of about 1-20 kD and is from about 1-100, 1-50, 1-30,1-25 or 15-25 nucleotides in length. In another aspect, theoligonucleotide has a molecular weight of about 5-10 kD. In one aspect,the alkylglycoside is tetradecyl-beta-D-maltoside.

In yet another aspect, the invention provides a method of increasing thebioavailability of a low molecular weight oligonucleotide in a subjectby administering the compound with an absorption increasing amount of analkylglycoside, thereby increasing the bioavailability of the compoundin the subject. In one aspect, the alkylglycoside istetradecyl-beta-D-maltoside.

In one aspect, the invention provides a method of increasing absorptionof a compound into the CSF of a subject having administered intranasallythe compound and an absorption increasing amount of a suitable nontoxic,nonionic alkyl glycoside having a hydrophobic alkyl group joined by alinkage to a hydrophilic saccharide.

In yet another aspect, the invention provides a pharmaceuticalcomposition having a suitable nontoxic, nonionic alkyl glycoside havinga hydrophobic alkyl group joined by a linkage to a hydrophilicsaccharide in combination with a mucosal delivery-enhancing agentselected from:

-   -   (a) an aggregation inhibitory agent;    -   (b) a charge-modifying agent;    -   (c) a pH control agent;    -   (d) a degradative enzyme inhibitory agent;    -   (e) a mucolytic or mucus clearing agent;    -   (f) a ciliostatic agent;    -   (g) a membrane penetration-enhancing agent selected from:        -   (i) a surfactant;        -   (ii) a bile salt;        -   (ii) a phospholipid additive, mixed micelle, liposome, or            carrier;        -   (iii) an alcohol;        -   (iv) an enamine;        -   (v) an NO donor compound;        -   (vi) a long-chain amphipathic molecule;        -   (vii) a small hydrophobic penetration enhancer;        -   (viii) sodium or a salicylic acid derivative;        -   (ix) a glycerol ester of acetoacetic acid;        -   (x) a cyclodextrin or beta-cyclodextrin derivative;        -   (xi) a medium-chain fatty acid;        -   (xii) a chelating agent;        -   (xiii) an amino acid or salt thereof;        -   (xiv) an N-acetylamino acid or salt thereof;        -   (xv) an enzyme degradative to a selected membrane component;        -   (ix) an inhibitor of fatty acid synthesis;        -   (x) an inhibitor of cholesterol synthesis; and        -   (xi) any combination of the membrane penetration enhancing            agents recited in (i)-(x);    -   (h) a modulatory agent of epithelial junction physiology;    -   (i) a vasodilator agent;    -   (j) a selective transport-enhancing agent; and    -   (k) a stabilizing delivery vehicle, carrier, mucoadhesive,        support or complex-forming species with which the compound is        effectively combined, associated, contained, encapsulated or        bound resulting in stabilization of the compound for enhanced        nasal mucosal delivery, wherein the formulation of the compound        with the intranasal delivery-enhancing agents provides for        increased bioavailability of the compound in a blood plasma of a        subject.

In one aspect, the invention provides a method of increasing absorptionof a low molecular weight compound into the circulatory system of asubject by administering, via the oral, ocular, nasal, nasolacrimal,inhalation or pulmonary, oral cavity (sublingual or Buccal cell) or CSFdelivery route (a) the compound; (b) an absorption increasing amount ofa suitable nontoxic, nonionic alkyl glycoside having a hydrophobic alkylgroup joined by a linkage to a hydrophilic saccharide; and (c) a mucosaldelivery-enhancing agent.

In one aspect, the invention provides a method of controlling caloricintake by administering a composition having a therapeutic effectiveamount of exendin-4, or related GLP-1 peptide, with an effective amountof Intravail alkyl saccharide.

In another aspect, the invention provides a method of controlling bloodglucose levels in a subject by administering to a subject a compositioncomprising a therapeutic effective amount of exendin-4, or related GLP-1peptide, with an effective amount of Intravail alkyl saccharide.

Still, in another aspect, the invention provides a controlled releasedosage composition comprising:

-   -   (a) a core comprising:        -   (i) at least one therapeutic agent or drug;        -   (ii) at least one alkyl glycoside and/or saccharide alkyl            ester; and    -   (b) at least one membrane coating surrounding the core, wherein        the coating is impermeable, permeable, semi-permeable or porous        and becomes more permeable upon sustained contact with contents        of the gastrointestinal tract.

In another embodiment, the invention provides a method of administeringan alkylglycoside composition by administering a therapeuticallyeffective amount of at least one alkyglycoside having an alkyl chainlength from about 12 to about 14 carbon atoms, at least one saccharidewith an antibacterial activity, and at least one therapeutic agent.

Still in another embodiment, the invention provides a composition havingat least one drug selected from the group consisting of insulin, PYY,Exendin-4 or other GLP-1 related peptide, human growth hormone,calcitonin, parathyroid hormone, truncated parathyroid hormone peptidessuch as PTH 1-34, EPO, interferon alpha, interferon beta, interferongamma, and GCSF and at least one alkyl saccharide having antibacterialactivity.

In one aspect, the invention provides an antibacterial alkyl saccharidecomposition, which includesn-Dodecyl-4-O-α-D-glucopyranosyl-β-D-glucopyranoside orn-tetradecyl-4-O-α-D-glucopyranosyl-β-D-glucopyranoside.

Yet, in another aspect, the invention provides an aqueous drugcomposition for transmucocal or transdermal administration having atleast one drug and at least one antibacterial agent in a concentrationfrom about 0.05% to about 0.5%.

In another aspect, the invention provides a fast-dispersing drugformulation containing a matrix material and an alkylsaccharide. Theformulation may have a Tmax substantially less than, and a first-passeffect substantially less than that observed for an equivalentformulation not containing an alkylsaccharide. In one embodiment, theformulation may contain about 0.1% to 10% alkylsaccharide, and exhibitsa Tmax substantially less than six hours and a first-pass effect of lessthan 40%. The alkylglycoside may be any suitable alykylglycoside and ina preferred aspect is dodecyl maltoside, tetradecyl maltoside, sucrosedodecanoate, or sucrose mono- and di-stearate. The formulation mayinclude a variety of different therapeutics, such as but not limited tomelatonin, raloxifene, olanzapene and diphenhydramine.

In another aspect, the invention provides a method for providing anextended absorption curve by attenuating the alkylsaccharideconcentration in drug formulation to balance gastric and buccaldelivery. For example, this is performed by providing a drug formulationincluding a matrix material and an alkylsaccharide having a Tmaxsubstantially less than, and a first-pass effect substantially less thanthat observed for an equivalent formulation not containing analkylsaccharide.

In one aspect, the invention provides a pharmaceutical compositionhaving a therapeutically effective amount of a bisphosphonate analog ora triptan analog in combination with an absorption increasing amount ofan alkylglycoside. In various embodiments, the bisphosphonate analog maybe etidronate, clodronate, tiludronate, pamidronate, neridronate,olpadronate, alendronate, ibandronate, risedronate, zoledronate, and/orpharmaceutically acceptable analogs thereof. In an exemplary embodiment,the bisphosphonate analog is alendronate or pharmaceutically acceptableanalog thereof. In various embodiments, the triptan analog may besumatriptan, rizatriptan, naratriptan, zolmitriptan, eletriptan,almotriptan, frovatriptan and/or pharmaceutically acceptable analogsthereof. In an exemplary embodiment, the triptan analog is sumatriptanor pharmaceutically acceptable analog thereof. In various embodiments,the alkylglycoside is tetradecyl-beta-D-maltoside.

In yet another aspect, the invention provides a method of increasing thebioavailability of a bisphosphonate analog or a triptan analog in asubject by administering the compound with an absorption increasingamount of an alkylglycoside, thereby increasing the bioavailability ofthe compound in the subject.

In still another aspect, the invention provides a composition includinga peptide, wherein the peptide includes a D-amino acid or a site forcyclization, or combination thereof, and at least one alkylsaccharide,wherein the alkylsaccharide provides increased enteral absorption of thepeptide.

In yet another aspect, the invention provides method of increasingenteral adsorption of a peptide in a biphasic manner. The methodincludes orally or nasally administering to a subject a compositioncomprising at least one peptide, wherein the peptide comprises a D-aminoacid or a site for cyclization, or combination thereof, and at least onealkylsaccharide, wherein the enteral absorption of the peptide isincreased and systemic serum levels of the peptide are increased in abiphasic manner.

In yet another aspect, the invention provides a method of increasing thebioavailability of a glucagon-like peptide-1 (GLP-1) analog in asubject. The method includes administering the analog with an absorptionincreasing amount of an alkylglycoside, thereby increasing thebioavailability of the analog in the subject.

In yet another aspect, the invention provides a pharmaceuticalcomposition including a glucagon-like peptide-1 (GLP-1) analog; and anabsorption increasing amount of an alkylglycoside.

In another aspect, the invention provides a pharmaceutical compositionhaving an opioid compound in combination with an absorption increasingamount of an alkylglycoside. In various embodiments, the opioid compoundis of structural Formula I as defined herein, or a pharmaceuticallyacceptable salt thereof. In embodiments, the opioid compound is acompound selected from those set forth in Table XXII. In variousembodiments, the alkylglycoside is dodecyl-beta-D-maltoside.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the intranasal percent bioavailabilitycompared to intravenous injection and the subject-to-subjectcoefficients of variation for MIACALCIN® (salmon calcitonin) with andwithout alkyl glycoside.

FIG. 2 is a graph showing the effect of intranasal administration ofinsulin/0.25% TDM (filled circles) and intranasal administration ofinsulin alone (open circles) in reducing blood glucose levels.

FIG. 3 is a graph showing the effect of intranasal (closed triangles)and intraperitoneal (IP) injection (closed circles) administration ofexendin-4/0.25% TDM and IP injection of saline alone, minus TDM (opencircles) in reducing blood glucose levels following intraperitoneal (IP)injection of glucose (i.e., in a so-called “glucose tolerance test”).

FIG. 4 is a graph showing the uptake of 1 mg mouse p-Leu-4]OB3 in 0.3%alkylglycoside tetradecyl-beta-D-maltoside (Intravail™ A3) by male SwissWebster Mice following administration by gavage.

FIG. 5 is a graph showing the uptake of sumatriptan in 0.5%alkylglycoside tetradecyl-beta-D-maltoside (Intravail™ A3) by caninesfor both oral and rectal administration.

FIG. 6 is a graph showing the uptake profile of 30 μg octreotide insodium acetate buffer after subcutaneous delivery to male Swiss Webstermice.

FIG. 7 is a graph showing the uptake profile of 30 μg octreotide in 0.5%Intravail™ after oral delivery to male Swiss Webster mice.

FIG. 8 is a graph showing the uptake profile of 30 μg octreotide in 1.5%Intravail™ after oral delivery to male Swiss Webster mice.

FIG. 9 is a graph showing the uptake profile of 30 μg octreotide in 3.0%Intravail™ after oral delivery to male Swiss Webster mice.

FIG. 10 is a graph showing blood glucose levels after oraladministration of an alkylglycoside composition including liraglutideand challenge with glucose.

FIG. 11 is a graph displaying a dose response curve.

FIG. 12 is a graph showing the percent of PE in the plasma (caninemodel) over time when delivered with alkylglycoside,n-dodecyl-beta-D-maltoside.

FIG. 13 is graph showing the percent of PE in the plasma (canine model)over time when delivered with the alkylglycoside, sucrosemonododecanoate.

FIG. 14 is a graph of the average plasma levels of patients nasallyadministered sumatriptan.

FIG. 15 is a graph of the average plasma levels of patients nasallyadministered sumatriptan.

DETAILED DESCRIPTION OF THE INVENTION

The present invention may be understood more readily by reference to thefollowing detailed description of specific embodiments and the Examplesincluded therein.

The present invention is based on the discovery that therapeuticcompositions comprising of least one drug and at least one surfactant,wherein the surfactant is comprised of at least one alkyl glycosideand/or at least one saccharide alkyl ester are stable, non-toxic,non-irritating, anti-bacterial compositions that increasebioavailability of the drug and have no observable adverse effects whenadministered to a subject.

A “therapeutic composition” can consist of an admixture with an organicor inorganic carrier or excipient, and can be compounded, for example,with the usual non-toxic, pharmaceutically acceptable carriers fortablets, pellets, capsules, suppositories, solutions, emulsions,suspensions, or other form suitable for use. The carriers, in additionto those disclosed above, can include glucose, lactose, mannose, gumacacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc,corn starch, keratin, colloidal silica, potato starch, urea, mediumchain length triglycerides, dextrans, and other carriers suitable foruse in manufacturing preparations, in solid, semisolid, or liquid form.In addition, auxiliary stabilizing, thickening or coloring agents can beused, for example a stabilizing dry agent such as triulose.

A “drug” is any therapeutic compound, or molecule, or therapeutic agent,or biologically active compound, including but not limited to nucleicacids, small molecules, proteins, polypeptides or peptides and the like.

The term “nucleic acids” or “oligonucleotide” also denotes DNA, cDNA,RNA, siRNA, RNAi, dsRNA and the like, which encode translated anduntranslated regions or inhibits translated or untranslated regions ofstructural genes encoding a peptide or protein or regulatory region. Forexample, a nucleic acid of the invention can include 5′ and 3′untranslated regulatory nucleotide sequences as well as translatedsequences associated with a structural gene. The term “nucleic acids” or“oligonucleotide” or grammatical equivalents as used herein, refers toat least two nucleotides covalently linked together.

Additionally, the term “oligonucleotide” refers to structures includingmodified portions such as modified sugar moieties, modified basemoieties or modified sugar linking moieties. These modified portionsfunction in a manner similar to natural bases, natural sugars andnatural phosphodiester linkages. Accordingly, oligonucleotides may havealtered base moieties, altered sugar moieties or altered inter-sugarlinkages. Modified linkages may be, for example, phosphoramide,phosphorothioate, phosphorodithioate, methyl phosphonate,phosphotriester, phosphoramidate, O-methylphophoroamidite linkages, orpeptide nucleic acid backbones and linkages. Other analogs may includeoligonucleotides with positive backbones, non-ionic backbones andnon-ribose backbones. The nucleic acid may be DNA, both genomic andcDNA, RNA or a hybrid, where the nucleic acid contains any combinationof deoxyribo- and ribo-nucleotides, and any combination of natural ormodified bases, including uracil, adenine, thymine, cytosine, guanine,inosine, xathanine, hypoxathanine, isocytosine, isoguanine, halogentatedbases and the like. Other modifications may include, for example, deazaor aza purines and pyrimidines used in place of natural purine andpyrimidine bases; pyrimidine bases having substituent groups at the 5-or 6-positions, purine bases having altered or replacement substituentgroups at the 2-, 6- or 8-positions, or sugars having substituent groupsat their 2′-position, substitutions for one or more of the hydrogenatoms of the sugar, or carbocyclic or acyclic sugars.

The term “antisense,” as used herein, refers to any compositioncontaining a nucleic acid sequence which is complementary to a specificnucleic acid sequence. The term “antisense strand” is used in referenceto a nucleic acid strand that is complementary to the “sense” strand.Antisense molecules may be produced by any method including synthesis ortranscription. Once introduced into a cell, the complementarynucleotides combine with natural sequences produced by the cell to formduplexes and to block either transcription or translation.

Antisense molecules include oligonucleotides comprising a singe-strandednucleic acid sequence (either RNA or DNA) capable of binding to targetreceptor or ligand mRNA (sense) or DNA (antisense) sequences. Theability to derive an antisense or a sense oligonucleotide, based upon acDNA sequence encoding a given protein. Antisense or senseoligonucleotides further comprise oligonucleotides having modifiedsugar-phosphodiester backbones and wherein such sugar linkages areresistant to endogenous nucleases. Such oligonucleotides with resistantsugar linkages are stable in vivo (i.e., capable of resisting enzymaticdegradation) but retain sequence specificity to be able to bind totarget nucleotide sequences.

RNAi is a phenomenon in which the introduction of dsRNA into a diverserange of organisms and cell types causes degradation of thecomplementary mRNA. In the cell, long dsRNAs are cleaved into short(e.g., 21-25 nucleotide) small interfering RNAs (siRNAs), by aribonuclease. The siRNAs subsequently assemble with protein componentsinto an RNA-induced silencing complex (RISC), unwinding in the process.The activated RISC then binds to complementary transcripts by basepairing interactions between the siRNA antisense strand and the mRNA.The bound mRNA is then cleaved and sequence specific degradation of mRNAresults in gene silencing. As used herein, “silencing” refers to amechanism by which cells shut down large sections of chromosomal DNAresulting in suppressing the expression of a particular gene. The RNAimachinery appears to have evolved to protect the genome from endogenoustransposable elements and from viral infections. Thus, RNAi can beinduced by introducing nucleic acid molecules complementary to thetarget mRNA to be degraded.

Other examples of sense or antisense oligonucleotides include thoseoligonucleotides which are covalently linked to organic moieties andother moieties that increase affinity of the oligonucleotide for atarget nucleic acid sequence, such as poly-(L-lysine). Further still,intercalating agents, such as ellipticine, and alkylating agents ormetal complexes may be attached to sense or antisense oligonucleotidesto modify binding specificities of the antisense or senseoligonucleotide for the target nucleotide sequence.

A peptide of the invention may be any medically or diagnostically usefulpeptide or protein of small to medium size (i.e., up to about 15 kD, 30kD, 40 kD, 50 kD, 60 kD, 70 kD, 80 kD, 90 kD, 100 kD, for example). Themechanisms of improved polypeptide absorption are described in U.S. Pat.No. 5,661,130 which is hereby incorporated by reference in its entirety.Invention compositions can be mixed with all such peptides, although thedegree to which the peptide benefits are improved may vary according tothe molecular weight and the physical and chemical properties of thepeptide, and the particular surfactant used. Examples of polypeptidesinclude vasopressin, vasopressin polypeptide analogs, desmopressin,glucagon, corticotropin (ACTH), gonadotropin, calcitonin, C-peptide ofinsulin, parathyroid hormone (PTH), growth hormone (HG), human growthhormone (hGH), growth hormone releasing hormone (GHRH), oxytocin,corticotropin releasing hormone (CRH), somatostatin or somatostatinpolypeptide analogs, gonadotropin agonist or gonadotrophin agonistpolypeptide analogs, human atrial natriuretic peptide (ANP), humanthyroxine releasing hormone (TRH), follicle stimulating hormone (FSH),prolactin, insulin, insulin like growth factor-I (IGF-I)somatomedin-C(SM-C), calcitonin, leptin and the leptin derived shortpeptide OB-3, melatonin, GLP-1 or Glucagon-like peptide-1 and analogsthereof, such as exenatide, albiglutide, taspoglutide, liraglutide andlixisenatide, GiP, neuropeptide pituitary adenylate cyclase, GM-1ganglioside, nerve growth factor (NGF), nafarelin, D-tryp6)-LHRH, FGF,VEGF antagonists, leuprolide, interferon (e.g., α,β,γ) low molecularweight heparin, PYY, LHRH antagonists, Keratinocyte Growth Factor (KGF),Glial-Derived Neurotrophic Factor (GDNF), ghrelin, and ghrelinantagonists. Further, in some aspects, the peptide or protein isselected from a growth factor, interleukin, polypeptide vaccine, enzyme,endorphin, glycoprotein, lipoprotein, or a polypeptide involved in theblood coagulation cascade.

Certain short peptides composed of approximately 8 to 10 D-amino acidsdesignated Allosteramers® produced by Allostera Pharma Inc., Quebec,Canada, have been shown to have an increased degree of oralbioavailability as well as extended length of time in the blood stream.Such D-amino acid-containing peptides are particularly well suited foruse with the present invention. Cyclization, as in cyclic PTH 1-31(Nemeth 2008), provides another way to reduce gastrointestinalhydrolysis. Thus, in various aspects, short peptides containingnon-naturally occurring structural modifications or amino acids are bestsuited to the present invention. Peptides comprising less than about 60,50, 40, 30, 20, 15 or 10 amino acids are contemplated.

Another example of a peptide containing D-amino acids is the D-Leu OB-3peptide, which is orally active when administered in combination withalkylglycosides, such as n-dodecyl-beta-D-maltoside.

Another peptide for use with the present invention is octreotide acetate(Sandostatin®). Octreotide is a cyclic octapeptide used foradministration by deep subcutaneous (intrafat) or intravenous injectionfor treatment of acromegaly, metastatic carcinoid tumors where itsuppresses or inhibits the severe diarrhea and flushing episodesassociated with the disease, and the treatment of the profuse waterydiarrhea associated with VIP-secreting tumors. Octreotide acetate isknown chemically as L-Cysteinamide,D-phenylalanyl-L-cysteinyl-L-phenylalanyl-D-tryptophyl-L-lysyl-L-threonyl-N-[2-hydroxy-1-(hydroxymethyl)propyl]-,cyclic (2→7)-disulfide; [R—(R*, R*)] acetate salt. It is a long-actingoctapeptide with pharmacologic actions mimicking those of the naturalhormone somatostatin and it contains both D-amino acids as well ascyclization, two properties that stabilize the molecule againstdestruction in the gastrointestinal tract. Octreotide is currently onlyadministered by injection, however as discussed herein, may besuccessfully delivered nasally or orally. Analogs of somatostatin havingaltered amino acyl sequences have also been prepared and are suitablefor use with the present invention.

In one aspect, the present invention provides oral administration ofoctreotide or octreotide analogs, such as but not limited topentetreotide Dicarba-Analog of Octreotide, or I-123 Tyr3-octreotidewith high bioavailability, circumventing the need and inconvenience ofmultiple daily and monthly injections and preventing needle stickinjuries and associated infections of healthcare providers and familymembers.

Other drugs or therapeutic compounds, molecules and/or agents includecyclic peptides, such as oxytocin, carbetocin, and demoxytocin,compounds or molecules of the central nervous system affectingneurotransmitters or neural ion channels (i.e., antidepressants(bupropion)), selective serotonin 2c receptor agonists, anti-seizureagents (topiramate, zonisamide), some dopamine antagonists, andcannabinoid-1 receptor antagonists (rimonabant)); leptin/insulin/centralnervous system pathway agents (i.e., leptin analogues, leptin transportand/or leptin receptor promoters, ciliary neurotrophic factor (Axokine),neuropeptide Y and agouti-related peptide antagonists,proopiomelanocortin, cocaine and amphetamine regulated transcriptpromoters, alpha-melanocyte-stimulating hormone analogues,melanocortin-4 receptor agonists, protein-tyrosine phosphatase-1Binhibitors, peroxisome proliferator activated receptor-gamma receptorantagonists, short-acting bromocriptine (ergoset), somatostatin agonists(octreotide), and adiponectin); gastrointestinal-neural pathway agents(i.e., agents that increase glucagon-like peptide-1 activity, such asexenatide (extendin-4), liraglutide, taspoglutide, albiglutide,lixisenatide and dipeptidyl peptidase IV inhibitors, protein YY3-36,ghrelin, ghrelin antagonists, amylin analogues (pramlintide)); andcompounds or molecules that may increase resting metabolic rate“selective” beta-3 stimulators/agonist, melanin concentrating hormoneantagonists, phytostanol analogues, functional oils, P57, amylaseinhibitors, growth hormone fragments, synthetic analogues ofdehydroepiandrosterone sulfate, antagonists of adipocyte11B-hydroxysteroid dehydrogenase type 1 activity,corticotropin-releasing hormone agonists, inhibitors of fatty acidsynthesis, carboxypeptidase inhibitors, gastrointestinal lipaseinhibitors (ATL962), melatonin, raloxifene, olanzapene anddiphenhydramine.

Other drugs or therapeutic compounds include osteoporosis drugs, such asbisphosphonate analogs. Bisphosphonate analogs, also known asdiphosphonates, are used clinically for the treatment of conditions suchas osteoporosis, osteitis deformans (Paget's disease of the bone), bonemetastasis (with or without hypercalcaemia), multiple myeloma,osteogenesis imperfecta and other conditions that feature bonefragility. The class of drugs inhibit osteoclast action and theresorption of bone. Examples of bisphosphonates to be admixed withalkylsaccharides for use in the compositions as described herein includeboth non-N-containing and N-containing bisphosphonate analogs. Exampleof non-N-containing bisphosphonates include etidronate (Didronel™),clodronate (Bonefos™, Loron™), tiludronate (Skelid™), andpharmaceutically acceptable analogs thereof. Examples of N-containingbisphosphonates include pamidronate (Aredia™), neridronate, olpadronate,alendronate (Fosamax™ or Fosamax-D™), ibandronate (Boniva™), risedronate(Actonel™), and zoledronate (Zometa™ or Reclast™), and pharmaceuticallyacceptable analogs thereof.

Other drugs or therapeutic compounds include drugs, such as triptananalogs. Triptan analogs are generally a family of tryptamine baseddrugs used for the treatment of migraines and headaches. Their action isattributed to their binding to serotonin receptors in nerve ending andin cranial blood vessels (causing their constriction) and subsequentinhibition of pro-inflammatory neuropeptide release. Examples oftriptans to be admixed with alkylsaccharides for use in the compositionsas described herein include sumatriptan (Imitrex™ and Imigran™),rizatriptan (Maxalt™), naratriptan (Amerge™ and Naramig™), zolmitriptan(Zomig™), eletriptan (Relpax™), almotriptan (Axert™ and Almogran™),frovatriptan (Frova™ and Migard™), and pharmaceutically acceptableanalogs thereof.

Additional drugs or therapeutic compounds include compounds, such asopioids. Opioids are psychoactive compounds that resemble morphine orother opiates in their pharmacological effects. Opioids generally workby binding to opioid receptors, which are found principally in thecentral and peripheral nervous system and the gastrointestinal tract. Asused herein, the term “opioid” refers to opiates, i.e., naturalalkaloids found in the resin of the opium poppy (Papaver somniferum), aswell as synthetic compounds.

In embodiments, an opioid compound is of structural Formula I, or apharmaceutically acceptable salt thereof:

-   -   wherein:    -   is a single or double bond;    -   R1 is selected from the group consisting of hydrogen, alkyl and        acyl;    -   R2 is alkyl or null;    -   R3 is selected from the group consisting of substituted or        unsubstituted alkyl, cycloalkylalkyl and alkenylalkyl;    -   R4 is hydrogen or hydroxyl or R4 may be taken together with R7        to form a carbocycle;    -   R5 is selected from the group consisting of hydrogen,        substituted or unsubstituted alkyl and substituted or        unsubstituted hydroxyalkyl;    -   R6 is hydrogen or —OR8;    -   R7 is hydrogen or R7 and R6 may be taken together to form a        carbonyl group; and    -   R8 is selected from the group consisting of hydrogen,        substituted or unsubstituted alkyl and acyl.

Various opioid compounds envisioned for use in the present invention areset forth in Table XXII.

TABLE XXII Opioid Compounds Structure Name

morphine

codeine

thebaine

oripavine

heroin

oxycodone

hydrocodone

naltrexone

naloxone

buprenorphine

nalbuphine

methylnaltrexone

Additional opioid compounds envisioned for use in the present inventioninclude 2,4-Dinitrophenylmorphine, 6-MDDM, chlomaltrexamine,desomorphine, dihydromorphine, hydromorphinol, methyldesorphine,N-Phenethylnormorphine, RAM-378, acetylpropionylmorphine, dihydroheroin,dibenzoylmorphine, dipropanoylmorphine, nicomorphine, 6-MAC,benzylmorphine, codeine methylbromide, dihydroheterocodeine,ethylmorphine, heterocodeine, pholcodine, myrophine,14-Cinnamoyloxycodeinone, 14-Ethoxymetopon, 14-Methoxymetopon, PPOM,7-Spiroindanyloxymorphone, acetylmorphone, codeinone, conorphone,codoxime, thebacon, hydrocodone, hydromorphone, metopon, morphinone,N-Phenethyl-14-Ethoxymetopon, oxycodone, oxymorphone, pentamorphone,semorphone, chloromorphide, 14-Hydroxydihydrocodeine,acetyldihydrocodeine, dihydrocodeine, nalbuphine, nicocodeine,nicodicodeine, oxymorphazone, 1-Iodomorphine, M6G, 6-MAM, norcodeine,normorphine, morphine-N-oxide, cyclorphan, DXA, levorphanol,levophenacylmorphan, levomethorphan, dextromethorphan, norlevorphanol,oxilorphan, phenomorphan, furethylnorlevorphanol, xorphanol,butorphanol, cyprodime, drotebanol, 7-PET, acetorphine, BU-48,buprenorphine, cyprenorphine, dihydroetorphine, etorphine,norbuprenorphine, 5′-Guanidinonaltrindole, diprenorphine, levallorphan,MNTX, nalfurafine, nalmefene, naloxazone, nalorphine, naltrexone,naltriben, naltrindole, 6β-Naltrexol-d4, pseudomorphine, naloxonazineand norbinaltorphimine.

As used herein, the terms below have the meanings indicated.

When ranges of values are disclosed, and the notation “from n1 . . . ton2” or “between n1 . . . and n2” is used, where n1 and n2 are thenumbers, then unless otherwise specified, this notation is intended toinclude the numbers themselves and the range between them. This rangemay be integral or continuous between and including the end values. Byway of example, the range “from 2 to 6 carbons” is intended to includetwo, three, four, five, and six carbons, since carbons come in integerunits. Compare, by way of example, the range “from 1 to 3 μM(micromolar),” which is intended to include 1 μM, 3 μM, and everythingin between to any number of significant figures (e.g., 1.255 μM, 2.1 μM,2.9999 μM, etc.). When n is set at 0 in the context of “0 carbon atoms”,it is intended to indicate a bond or null.

The term “about,” as used herein, is intended to qualify the numericalvalues which it modifies, denoting such a value as variable within amargin of error. When no particular margin of error, such as a standarddeviation to a mean value given in a chart or table of data, is recited,the term “about” should be understood to mean that range which wouldencompass the recited value and the range which would be included byrounding up or down to that figure as well, taking into accountsignificant figures.

The term “acyl,” as used herein, alone or in combination, refers to acarbonyl attached to an alkenyl, alkyl, aryl, cycloalkyl, heteroaryl,heterocycle, or any other moiety where the atom attached to the carbonylis carbon. An “acetyl” group refers to a—C(O)CH3 group. An“alkylcarbonyl” or “alkanoyl” group refers to an alkyl group attached tothe parent molecular moiety through a carbonyl group. Examples of suchgroups include methylcarbonyl and ethylcarbonyl. Examples of acyl groupsinclude formyl, alkanoyl and aroyl.

The term “alkenyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain hydrocarbon group having one or moredouble bonds and containing from 2 to 20 carbon atoms. In certainembodiments, said alkenyl will comprise from 2 to 6 carbon atoms. Theterm “alkenylene” refers to a carbon-carbon double bond system attachedat two or more positions such as ethenylene [(—CH═CH—), (—C::C—)].Examples of suitable alkenyl groups include ethenyl, propenyl,2-methylpropenyl, 1,4-butadienyl and the like. Unless otherwisespecified, the term “alkenyl” may include “alkenylene” groups.

The term “alkoxy,” as used herein, alone or in combination, refers to analkyl ether group, wherein the term alkyl is as defined below. Examplesof suitable alkyl ether groups include methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, iso-butoxy, sec-butoxy, tert-butoxy, and the like.

The term “alkyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain alkyl group containing from 1 to 20carbon atoms. In certain embodiments, said alkyl will comprise from 1 to10 carbon atoms. In further embodiments, said alkyl will comprise from 1to 6 carbon atoms. Alkyl groups may be optionally substituted as definedherein. Examples of alkyl groups include methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl,hexyl, octyl, noyl and the like. The term “alkylene,” as used herein,alone or in combination, refers to a saturated aliphatic group derivedfrom a straight or branched chain saturated hydrocarbon attached at twoor more positions, such as methylene (—CH2—). Unless otherwisespecified, the term “alkyl” may include “alkylene” groups.

The term “alkylamino,” as used herein, alone or in combination, refersto an alkyl group attached to the parent molecular moiety through anamino group. Suitable alkylamino groups may be mono- or dialkylated,forming groups such as, for example, N-methylamino, N-ethylamino,N,N-dimethylamino, N,N-ethylmethylamino and the like.

The term “alkylidene,” as used herein, alone or in combination, refersto an alkenyl group in which one carbon atom of the carbon-carbon doublebond belongs to the moiety to which the alkenyl group is attached.

The term “alkylthio,” as used herein, alone or in combination, refers toan alkyl thioether (R—S—) group wherein the term alkyl is as definedabove and wherein the sulfur may be singly or doubly oxidized. Examplesof suitable alkyl thioether groups include methylthio, ethylthio,n-propylthio, isopropylthio, n-butylthio, iso-butylthio, sec-butylthio,tert-butylthio, methanesulfonyl, ethanesulfinyl, and the like.

The term “alkynyl,” as used herein, alone or in combination, refers to astraight-chain or branched-chain hydrocarbon group having one or moretriple bonds and containing from 2 to 20 carbon atoms. In certainembodiments, said alkynyl comprises from 2 to 6 carbon atoms. In furtherembodiments, said alkynyl comprises from 2 to 4 carbon atoms. The term“alkynylene” refers to a carbon-carbon triple bond attached at twopositions such as ethynylene (—C:::C—, —C≡C—). Examples of alkynylgroups include ethynyl, propynyl, hydroxypropynyl, butyn-1-yl,butyn-2-yl, pentyn-1-yl, 3-methylbutyn-1-yl, hexyn-2-yl, and the like.Unless otherwise specified, the term “alkynyl” may include “alkynylene”groups.

The terms “amido” and “carbamoyl,” as used herein, alone or incombination, refer to an amino group as described below attached to theparent molecular moiety through a carbonyl group, or vice versa. Theterm “C-amido” as used herein, alone or in combination, refers to a—C(═O)—NR2 group with R as defined herein. The term “N-amido” as usedherein, alone or in combination, refers to a RC(═O)NH— group, with R asdefined herein. The term “acylamino” as used herein, alone or incombination, embraces an acyl group attached to the parent moietythrough an amino group. An example of an “acylamino” group isacetylamino (CH3C(O)NH—).

The term “amino,” as used herein, alone or in combination, refers to—NRR′, wherein R and R′ are independently selected from the groupconsisting of hydrogen, alkyl, acyl, heteroalkyl, aryl, cycloalkyl,heteroaryl, and heterocycloalkyl, any of which may themselves beoptionally substituted. Additionally, R and R′ may combine to formheterocycloalkyl, either of which may be optionally substituted. Theterm “aryl,” as used herein, alone or in combination, means acarbocyclic aromatic system containing one, two or three rings whereinsuch polycyclic ring systems are fused together. The term “aryl”embraces aromatic groups such as phenyl, naphthyl, anthracenyl, andphenanthryl.

The term “arylalkenyl” or “aralkenyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkenyl group.

The term “arylalkoxy” or “aralkoxy,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkoxy group.

The term “arylalkyl” or “aralkyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkyl group.

The term “arylalkynyl” or “aralkynyl,” as used herein, alone or incombination, refers to an aryl group attached to the parent molecularmoiety through an alkynyl group.

The term “arylalkanoyl” or “aralkanoyl” or “aroyl,” as used herein,alone or in combination, refers to an acyl group derived from anaryl-substituted alkanecarboxylic acid such as benzoyl, napthoyl,phenylacetyl, 3-phenylpropionyl (hydrocinnamoyl), 4-phenylbutyryl,(2-naphthyl)acetyl, 4-chlorohydrocinnamoyl, and the like.

The term aryloxy as used herein, alone or in combination, refers to anaryl group attached to the parent molecular moiety through an oxy.

The terms “benzo” and “benz,” as used herein, alone or in combination,refer to the divalent group C₆H₄═ derived from benzene. Examples includebenzothiophene and benzimidazole.

The term “carbamate,” as used herein, alone or in combination, refers toan ester of carbamic acid (—NHCOO—) which may be attached to the parentmolecular moiety from either the nitrogen or acid end, and which may beoptionally substituted as defined herein.

The term “O-carbamyl” as used herein, alone or in combination, refers toa —OC(O)NRR′, group-with R and R′ as defined herein.

The term “N-carbamyl” as used herein, alone or in combination, refers toa ROC(O)NR′— group, with R and R′ as defined herein.

The term “carbonyl,” as used herein, when alone includes formyl [—C(O)H]and in combination is a —C(O)— group.

The term “carboxyl” or “carboxy,” as used herein, refers to —C(O)OH orthe corresponding “carboxylate” anion, such as is in a carboxylic acidsalt. An “O-carboxy” group refers to a RC(O)O— group, where R is asdefined herein. A “C-carboxy” group refers to a —C(O)OR groups where Ris as defined herein.

The term “cyano,” as used herein, alone or in combination, refers to—CN.

The term “cycloalkyl,” or, alternatively, “carbocycle,” as used herein,alone or in combination, refers to a saturated or partially saturatedmonocyclic, bicyclic or tricyclic alkyl group wherein each cyclic moietycontains from 3 to 12 carbon atom ring members and which may optionallybe a benzo fused ring system which is optionally substituted as definedherein. In certain embodiments, said cycloalkyl will comprise from 5 to7 carbon atoms. Examples of such cycloalkyl groups include cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, tetrahydronapthyl,indanyl, octahydronaphthyl, 2,3-dihydro-1H-indenyl, adamantyl and thelike. “Bicyclic” and “tricyclic” as used herein are intended to includeboth fused ring systems, such as decahydronaphthalene,octahydronaphthalene as well as the multicyclic (multicentered)saturated or partially unsaturated type. The latter type of isomer isexemplified in general by, bicyclo[1,1,1]pentane, camphor, adamantane,and bicyclo[3,2,1]octane.

The term “ester,” as used herein, alone or in combination, refers to acarboxy group bridging two moieties linked at carbon atoms.

The term “ether,” as used herein, alone or in combination, refers to anoxy group bridging two moieties linked at carbon atoms.

The term “halo,” or “halogen,” as used herein, alone or in combination,refers to fluorine, chlorine, bromine, or iodine.

The term “haloalkoxy,” as used herein, alone or in combination, refersto a haloalkyl group attached to the parent molecular moiety through anoxygen atom.

The term “haloalkyl,” as used herein, alone or in combination, refers toan alkyl group having the meaning as defined above wherein one or morehydrogens are replaced with a halogen. Specifically embraced aremonohaloalkyl, dihaloalkyl and polyhaloalkyl groups. A monohaloalkylgroup, for one example, may have an iodo, bromo, chloro or fluoro atomwithin the group. Dihalo and polyhaloalkyl groups may have two or moreof the same halo atoms or a combination of different halo groups.Examples of haloalkyl groups include fluoromethyl, difluoromethyl,trifluoromethyl, chloromethyl, dichloromethyl, trichloromethyl,pentafluoroethyl, heptafluoropropyl, difluorochloromethyl,dichlorofluoromethyl, difluoroethyl, difluoropropyl, dichloroethyl anddichloropropyl. “Haloalkylene” refers to a haloalkyl group attached attwo or more positions. Examples include fluoromethylene (—CFH—),difluoromethylene (—CF₂—), chloromethylene (—CHCl—) and the like.

The term “heteroalkyl,” as used herein, alone or in combination, refersto a stable straight or branched chain, or cyclic hydrocarbon group, orcombinations thereof, fully saturated or containing from 1 to 3 degreesof unsaturation, consisting of the stated number of carbon atoms andfrom one to three heteroatoms selected from the group consisting of O,N, and S, and wherein the nitrogen and sulfur atoms may optionally beoxidized and the nitrogen heteroatom may optionally be quatemized. Theheteroatom(s) O, N and S may be placed at any interior position of theheteroalkyl group. Up to two heteroatoms may be consecutive, such as,for example, —CH₂—NH—OCH₃.

The term “heteroaryl,” as used herein, alone or in combination, refersto a 3 to 7 membered unsaturated heteromonocyclic ring, or a fusedmonocyclic, bicyclic, or tricyclic ring system in which at least one ofthe fused rings is aromatic, which contains at least one atom selectedfrom the group consisting of O, S, and N. In certain embodiments, saidheteroaryl will comprise from 5 to 7 carbon atoms. The term alsoembraces fused polycyclic groups wherein heterocyclic rings are fusedwith aryl rings, wherein heteroaryl rings are fused with otherheteroaryl rings, wherein heteroaryl rings are fused withheterocycloalkyl rings, or wherein heteroaryl rings are fused withcycloalkyl rings. Examples of heteroaryl groups include pyrrolyl,pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl,pyridazinyl, triazolyl, pyranyl, furyl, thienyl, oxazolyl, isoxazolyl,oxadiazolyl, thiazolyl, thiadiazolyl, isothiazolyl, indolyl, isoindolyl,indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, quinoxalinyl,quinazolinyl, indazolyl, benzotriazolyl, benzodioxolyl, benzopyranyl,benzoxazolyl, benzoxadiazolyl, benzothiazolyl, benzothiadiazolyl,benzofuryl, benzothienyl, chromonyl, coumarinyl, benzopyranyl,tetrahydroquinolinyl, tetrazolopyridazinyl, tetrahydroisoquinolinyl,thienopyridinyl, furopyridinyl, pyrrolopyridinyl and the like. Exemplarytricyclic heterocyclic groups include carbazolyl, benzidolyl,phenanthrolinyl, dibenzofuranyl, acridinyl, phenanthridinyl, xanthenyland the like.

The terms “heterocycloalkyl” and, interchangeably, “heterocycle,” asused herein, alone or in combination, each refer to a saturated,partially unsaturated, or fully unsaturated monocyclic, bicyclic, ortricyclic heterocyclic group containing at least one heteroatom as aring member, wherein each said heteroatom may be independently selectedfrom the group consisting of nitrogen, oxygen, and sulfur In certainembodiments, said hetercycloalkyl will comprise from 1 to 4 heteroatomsas ring members. In further embodiments, said hetercycloalkyl willcomprise from 1 to 2 heteroatoms as ring members. In certainembodiments, said hetercycloalkyl will comprise from 3 to 8 ring membersin each ring. In further embodiments, said hetercycloalkyl will comprisefrom 3 to 7 ring members in each ring. In yet further embodiments, saidhetercycloalkyl will comprise from 5 to 6 ring members in each ring.“Heterocycloalkyl” and “heterocycle” are intended to include sulfones,sulfoxides, N-oxides of tertiary nitrogen ring members, and carbocyclicfused and benzo fused ring systems; additionally, both terms alsoinclude systems where a heterocycle ring is fused to an aryl group, asdefined herein, or an additional heterocycle group. Examples ofheterocycle groups include aziridinyl, azetidinyl, 1,3-benzodioxolyl,dihydroisoindolyl, dihydroisoquinolinyl, dihydrocinnolinyl,dihydrobenzodioxinyl, dihydro[1,3]oxazolo[4,5-b]pyridinyl,benzothiazolyl, dihydroindolyl, dihy-dropyridinyl, 1,3-dioxanyl,1,4-dioxanyl, 1,3-dioxolanyl, isoindolinyl, morpholinyl, piperazinyl,pyrrolidinyl, tetrahydropyridinyl, piperidinyl, thiomorpholinyl, and thelike. The heterocycle groups may be optionally substituted unlessspecifically prohibited.

The term “hydrazinyl” as used herein, alone or in combination, refers totwo amino groups joined by a single bond, i.e., —N—N—.

The term “hydroxy,” as used herein, alone or in combination, refers to—OH.

The term “hydroxyalkyl,” as used herein, alone or in combination, refersto a hydroxy group attached to the parent molecular moiety through analkyl group.

The term “imino,” as used herein, alone or in combination, refers to═N—.

The term “iminohydroxy,” as used herein, alone or in combination, refersto ═N(OH) and ═N—O—.

The phrase “in the main chain” refers to the longest contiguous oradjacent chain of carbon atoms starting at the point of attachment of agroup to the compounds of any one of the formulas disclosed herein.

The term “isocyanato” refers to a —NCO group.

The term “isothiocyanato” refers to a —NCS group.

The phrase “linear chain of atoms” refers to the longest straight chainof atoms independently selected from carbon, nitrogen, oxygen andsulfur.

The term “lower,” as used herein, alone or in a combination, where nototherwise specifically defined, means containing from 1 to and including6 carbon atoms.

The term “lower aryl,” as used herein, alone or in combination, meansphenyl or naphthyl, which may be optionally substituted as provided.

The term “lower heteroaryl,” as used herein, alone or in combination,means either 1) monocyclic heteroaryl comprising five or six ringmembers, of which between one and four said members may be heteroatomsselected from the group consisting of O, S, and N, or 2) bicyclicheteroaryl, wherein each of the fused rings comprises five or six ringmembers, comprising between them one to four heteroatoms selected fromthe group consisting of O, S, and N.

The term “lower cycloalkyl,” as used herein, alone or in combination,means a monocyclic cycloalkyl having between three and six ring members.Lower cycloalkyls may be unsaturated. Examples of lower cycloalkylinclude cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term “lower heterocycloalkyl,” as used herein, alone or incombination, means a monocyclic heterocycloalkyl having between threeand six ring members, of which between one and four may be heteroatomsselected from the group consisting of O, S, and N. Examples of lowerheterocycloalkyls include pyrrolidinyl, imidazolidinyl, pyrazolidinyl,piperidinyl, piperazinyl, and morpholinyl. Lower heterocycloalkyls maybe unsaturated.

The term “lower amino,” as used herein, alone or in combination, refersto —NRR′, wherein R and R′ are independently selected from the groupconsisting of hydrogen, lower alkyl, and lower heteroalkyl, any of whichmay be optionally substituted. Additionally, the R and R′ of a loweramino group may combine to form a five- or six-memberedheterocycloalkyl, either of which may be optionally substituted.

The term “mercaptyl” as used herein, alone or in combination, refers toan RS— group, where R is as defined herein.

The term “nitro,” as used herein, alone or in combination, refers to—NO₂.

The terms “oxy” or “oxa,” as used herein, alone or in combination, referto —O—.

The term “oxo,” as used herein, alone or in combination, refers to ═O.

The term “perhaloalkoxy” refers to an alkoxy group where all of thehydrogen atoms are replaced by halogen atoms.

The term “perhaloalkyl” as used herein, alone or in combination, refersto an alkyl group where all of the hydrogen atoms are replaced byhalogen atoms.

The terms “sulfonate,” “sulfonic acid,” and “sulfonic,” as used herein,alone or in combination, refer to the —SO₃H group and its anion as thesulfonic acid is used in salt formation.

The term “sulfanyl,” as used herein, alone or in combination, refers to—S—.

The term “sulfinyl,” as used herein, alone or in combination, refers to—S(O)—.

The term “sulfonyl,” as used herein, alone or in combination, refers to—S(O)₂—.

The term “N-sulfonamido” refers to a RS(═O)₂NR′— group with R and R′ asdefined herein.

The term “S-sulfonamido” refers to a —S(═O)₂NRR′, group, with R and R′as defined herein.

The terms “thia” and “thio,” as used herein, alone or in combination,refer to a —S— group or an ether wherein the oxygen is replaced withsulfur. The oxidized derivatives of the thio group, namely sulfinyl andsulfonyl, are included in the definition of thia and thio.

The term “thiol,” as used herein, alone or in combination, refers to an—SH group.

The term “thiocarbonyl,” as used herein, when alone includes thioformyl—C(S)H and in combination is a —C(S)— group.

The term “N-thiocarbamyl” refers to an ROC(S)NR′— group, with R and R′as defined herein.

The term “O-thiocarbamyl” refers to a —OC(S)NRR′, group with R and R′ asdefined herein.

The term “thiocyanato” refers to a —CNS group.

The term “trihalomethanesulfonamido” refers to a X₃CS(O)₂NR— group withX is a halogen and R as defined herein.

The term “trihalomethanesulfonyl” refers to a X₃CS(O)₂— group where X isa halogen.

The term “trihalomethoxy” refers to a X₃CO— group where X is a halogen.

The term “trisubstituted silyl,” as used herein, alone or incombination, refers to a silicone group substituted at its three freevalences with groups as listed herein under the definition ofsubstituted amino. Examples include trimethysilyl,tert-butyldimethylsilyl, triphenylsilyl and the like.

Any definition herein may be used in combination with any otherdefinition to describe a composite structural group. By convention, thetrailing element of any such definition is that which attaches to theparent moiety. For example, the composite group alkylamido wouldrepresent an alkyl group attached to the parent molecule through anamido group, and the term alkoxyalkyl would represent an alkoxy groupattached to the parent molecule through an alkyl group.

When a group is defined to be “null,” what is meant is that said groupis absent.

The term “optionally substituted” means the anteceding group may besubstituted or unsubstituted. When substituted, the substituents of an“optionally substituted” group may include, without limitation, one ormore substituents independently selected from the following groups or aparticular designated set of groups, alone or in combination: loweralkyl, lower alkenyl, lower alkynyl, lower alkanoyl, lower heteroalkyl,lower heterocycloalkyl, lower haloalkyl, lower haloalkenyl, lowerhaloalkynyl, lower perhaloalkyl, lower perhaloalkoxy, lower cycloalkyl,phenyl, aryl, aryloxy, lower alkoxy, lower haloalkoxy, oxo, loweracyloxy, carbonyl, carboxyl, lower alkylcarbonyl, lower carboxyester,lower carboxamido, cyano, hydrogen, halogen, hydroxy, amino, loweralkylamino, arylamino, amido, nitro, thiol, lower alkylthio, lowerhaloalkylthio, lower perhaloalkylthio, arylthio, sulfonate, sulfonicacid, trisubstituted silyl, N₃, SH, SCH₃, C(O)CH₃, CO₂CH₃, CO₂H,pyridinyl, thiophene, furanyl, lower carbamate, and lower urea. Twosubstituents may be joined together to form a fused five-, six-, orseven-membered carbocyclic or heterocyclic ring consisting of zero tothree heteroatoms, for example forming methylenedioxy or ethylenedioxy.An optionally substituted group may be unsubstituted (e.g., —CH₂CH₃),fully substituted (e.g., —CF₂CF₃), monosubstituted (e.g., —CH₂CH₂F) orsubstituted at a level anywhere in-between fully substituted andmonosubstituted (e.g., —CH₂CF₃). Where substituents are recited withoutqualification as to substitution, both substituted and unsubstitutedforms are encompassed. Where a substituent is qualified as“substituted,” the substituted form is specifically intended.Additionally, different sets of optional substituents to a particularmoiety may be defined as needed; in these cases, the optionalsubstitution will be as defined, often immediately following the phrase,“optionally substituted with.”

The term R or the term R′, appearing by itself and without a numberdesignation, unless otherwise defined, refers to a moiety selected fromthe group consisting of hydrogen, alkyl, cycloalkyl, heteroalkyl, aryl,heteroaryl and heterocycloalkyl, any of which may be optionallysubstituted. Such R and R′ groups should be understood to be optionallysubstituted as defined herein. Whether an R group has a numberdesignation or not, every R group, including R, R′ and R^(n) where n=(1,2, 3, . . . n), every substituent, and every term should be understoodto be independent of every other in terms of selection from a group.Should any variable, substituent, or term (e.g., aryl, heterocycle, R,etc.) occur more than one time in a formula or generic structure, itsdefinition at each occurrence is independent of the definition at everyother occurrence. Those of skill in the art will further recognize thatcertain groups may be attached to a parent molecule or may occupy aposition in a chain of elements from either end as written. Thus, by wayof example only, an unsymmetrical group such as —C(O)N(R)— may beattached to the parent moiety at either the carbon or the nitrogen.

Asymmetric centers exist in the compounds disclosed herein. Thesecenters are designated by the symbols “R” or “S,” depending on theconfiguration of substituents around the chiral carbon atom. It shouldbe understood that the invention encompasses all stereochemical isomericforms, including diastereomeric, enantiomeric, and epimeric forms, aswell as d-isomers and 1-isomers, and mixtures thereof. Individualstereoisomers of compounds can be prepared synthetically fromcommercially available starting materials which contain chiral centersor by preparation of mixtures of enantiomeric products followed byseparation such as conversion to a mixture of diastereomers followed byseparation or recrystallization, chromatographic techniques, directseparation of enantiomers on chiral chromatographic columns, or anyother appropriate method known in the art. Starting compounds ofparticular stereochemistry are either commercially available or can bemade and resolved by techniques known in the art. Additionally, thecompounds disclosed herein may exist as geometric isomers. The presentinvention includes all cis, trans, syn, anti, entgegen (E), and zusammen(Z) isomers as well as the appropriate mixtures thereof. Additionally,compounds may exist as tautomers; all tautomeric isomers are provided bythis invention. Additionally, the compounds disclosed herein can existin unsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. In general, the solvatedforms are considered equivalent to the unsolvated forms.

The therapeutic composition of the invention includes a drug and a drugabsorption enhancing agent, for example, a surfactant. The term“surfactant” is any surface active agent that modifies interfacialtension of water. Typically, surfactants have one lipophilic and onehydrophilic group in the molecule. Broadly, the group includes soaps,detergents, emulsifiers, dispersing and wetting agents, and severalgroups of antiseptics. More specifically, surfactants includestearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionicacid, lecithin, benzalkonium chloride, benzethonium chloride andglycerin monostearate; and hydrophilic polymers such as polyvinylalcohol, polyvinylpyrrolidone, carboxymethylcellulose sodium,methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose andhydroxypropylcellulose.

Preferably, the surfactant of the invention consists of at least onesuitable alkyl glycoside. As used herein, “alkyl glycoside” refers toany sugar joined by a linkage to any hydrophobic alkyl, as is known inthe art. Any “suitable” alkyl glycoside means one that fulfills thelimiting characteristics of the invention, i.e., that the alkylglycoside be nontoxic and nonionic, and that it increases the absorptionof a compound when it is administered with the compound via the ocular,nasal, nasolacrimal, inhalation or pulmonary, oral cavity (sublingual orBuccal cell), or CSF delivery route. Suitable compounds can bedetermined using the methods set forth herein.

Alkyl glycosides of the invention can be synthesized by knownprocedures, i.e., chemically, as described, e.g., in Rosevear et al.,Biochemistry 19:4108-4115 (1980) or Koeltzow and Urfer, J. Am. Oil Chem.Soc., 61:1651-1655 (1984), U.S. Pat. Nos. 3,219,656 and 3,839,318 orenzymatically, as described, e.g., in Li et al., J. Biol. Chem.,266:10723-10726 (1991) or Gopalan et al., J. Biol. Chem. 267:9629-9638(1992).

Alkyl glycosides of the present invention can include, but are notlimited to: alkyl glycosides, such as octyl-, nonyl-, decyl-, undecyl-,dodecyl-, tridecyl-, tetradecyl-, pentadecyl-,hexadecyl-, heptadecyl-,and octadecyl-α- or β-D-maltoside, -glucoside or -sucroside (synthesizedaccording to Koeltzow and Urfer; Anatrace Inc., Maumee, Ohio;Calbiochem, San Diego, Calif.; Fluka Chemie, Switzerland); alkylthiomaltosides, such as heptyl, octyl, dodecyl-, tridecyl-, andtetradecyl-β-D-thiomaltoside (synthesized according to Defaye, J. andPederson, C., “Hydrogen Fluoride, Solvent and Reagent for CarbohydrateConversion Technology” in Carbohydrates as Organic Raw Materials,247-265 (F. W. Lichtenthaler, ed.) VCH Publishers, New York (1991);Ferenci, T., J. Bacteriol, 144:7-11 (1980)); alkyl thioglucosides, suchas heptyl- or octyl 1-thio α- or β-D-glucopyranoside (Anatrace, Inc.,Maumee, Ohio; see Saito, S. and Tsuchiya, T. Chem. Pharm. Bull.33:503-508 (1985)); alkyl thiosucroses (synthesized according to, forexample, Binder, T. P. and Robyt, J. F., Carbohydr. Res. 140:9-20(1985)); alkyl maltotriosides (synthesized according to Koeltzow andUrfer); long chain aliphatic carbonic acid amides of sucroseβ-amino-alkyl ethers; (synthesized according to Austrian Patent 382,381(1987); Chem. Abstr., 108:114719 (1988) and Gruber and Greber pp.95-116); derivatives of palatinose and isomaltamine linked by amidelinkage to an alkyl chain (synthesized according to Kunz, M.,“Sucrose-based Hydrophilic Building Blocks as Intermediates for theSynthesis of Surfactants and Polymers” in Carbohydrates as Organic RawMaterials, 127-153); derivatives of isomaltamine linked by urea to analkyl chain (synthesized according to Kunz); long chain aliphaticcarbonic acid ureides of sucrose β-amino-alkyl ethers (synthesizedaccording to Gruber and Greber, pp. 95-116); and long chain aliphaticcarbonic acid amides of sucrose β-amino-alkyl ethers (synthesizedaccording to Austrian Patent 382,381 (1987), Chem. Abstr., 108:114719(1988) and Gruber and Greber, pp. 95-116).

Surfactants of the invention consisting of an alkyl glycoside and/or asucrose ester have characteristic hydrophile-lipophile balance (HLB)numbers, which can be calculated or determined empirically (Schick, M.J. Nonionic Surfactants, p. 607 (New York: Marcel Dekker, Inc. (1967)).The HLB number is a direct reflection of the hydrophilic character ofthe surfactant, i.e., the larger the HLB number, the more hydrophilicthe compound. HLB numbers can be calculated by the formula: (20 times MWhydrophilic component)/(MW hydrophobic component+MW hydrophiliccomponent), where MW=molecular weight (Rosen, M. J., Surfactants andInterfacial Phenomena, pp. 242-245, John Wiley, New York (1978)). TheHLB number is a direct expression of the hydrophilic character of thesurfactant, i.e., the larger the HLB number, the more hydrophilic thecompound. A preferred surfactant has an HLB number of from about 10 to20 and an even more preferred range of from about 11 to 15.

As described above, the hydrophobic alkyl can thus be chosen of anydesired size, depending on the hydrophobicity desired and thehydrophilicity of the saccharide moiety. For example, one preferredrange of alkyl chains is from about 9 to about 24 carbon atoms. An evenmore preferred range is from about 9 to about 16 or about 14 carbonatoms. Similarly, some preferred glycosides include maltose, sucrose,and glucose linked by glycosidic linkage to an alkyl chain of 9, 10, 12,13, 14, 16, 18, 20, 22, or 24 carbon atoms, e.g., nonyl-, decyl-,dodecyl- and tetradecyl sucroside, glucoside, and maltoside, etc. Thesecompositions are nontoxic, since they are degraded to an alcohol and anoligosaccharide, and amphipathic.

The surfactants of the invention can also include a saccharide. As useherein, a “saccharide” is inclusive of monosaccharides, oligosaccharidesor polysaccharides in straight chain or ring forms, or a combinationthereof to form a saccharide chain. Oligosaccharides are saccharideshaving two or more monosaccharide residues. The saccharide can bechosen, for example, from any currently commercially availablesaccharide species or can be synthesized. Some examples of the manypossible saccharides to use include glucose, maltose, maltotriose,maltotetraose, sucrose and trehalose. Preferable saccharides includemaltose, sucrose and glucose.

The surfactants of the invention can likewise consist of a sucroseester. As used herein, “sucrose esters” are sucrose esters of fattyacids and is a complex of sucrose and fatty acid. Sucrose esters cantake many forms because of the eight hydroxyl groups in sucroseavailable for reaction and the many fatty acid groups, from acetate onup to larger, more bulky fatty acids that can be reacted with sucrose.This flexibility means that many products and functionalities can betailored, based on the fatty acid moiety used. Sucrose esters have foodand non-food uses, especially as surfactants and emulsifiers, withgrowing applications in pharmaceuticals, cosmetics, detergents and foodadditives. They are biodegradable, non-toxic and mild to the skin.

The surfactants of the invention have a hydrophobic alkyl group linkedto a hydrophilic saccharide. The linkage between the hydrophobic alkylgroup and the hydrophilic saccharide can include, among otherpossibilities, a glycosidic, thioglycosidic (Horton), amide(Carbohydrates as Organic Raw Materials, F. W. Lichtenthaler ed., VCHPublishers, New York, 1991), ureide (Austrian Pat. 386,414 (1988); Chem.Abstr. 110:137536p (1989); see Gruber, H. and Greber, G., “ReactiveSucrose Derivatives” in Carbohydrates as Organic Raw Materials, pp.95-116) or ester linkage (Sugar Esters: Preparation and Application, J.C. Colbert ed., (Noyes Data Corp., New Jersey), (1974)). Further,preferred glycosides can include maltose, sucrose, and glucose linked byglycosidic linkage to an alkyl chain of about 9-16 carbon atoms, e.g.,nonyl-, decyl-, dodecyl- and tetradecyl sucroside, glucoside, andmaltoside. Again, these compositions are amphipathic and nontoxic,because they degrade to an alcohol and an oligosaccharide.

The above examples are illustrative of the types of glycosides to beused in the methods claimed herein, but the list is not exhaustive.Derivatives of the above compounds which fit the criteria of the claimsshould also be considered when choosing a glycoside. All of thecompounds can be screened for efficacy following the methods taughtherein and in the examples.

The compositions of the present invention can be administered in aformat selected from the group consisting of a tablet, a capsule, asuppository, a drop, a spray, an aerosol and a sustained release ordelayed burst format. The spray and the aerosol can be achieved throughuse of an appropriate dispenser. The sustained release format can be anocular insert, erodible microparticulates, swelling mucoadhesiveparticulates, pH sensitive microparticulates, nanoparticles/latexsystems, ion-exchange resins and other polymeric gels and implants(Ocusert, Alza Corp., California; Joshi, A., S. Ping and K. J.Himmelstein, Patent Application WO 91/19481). These systems maintainprolonged drug contact with the absorptive surface preventing washoutand nonproductive drug loss. The prolonged drug contact is non-toxic tothe skin and mucosal surfaces.

The surfactant compositions of the invention are stable. For example,Baudys et al. in U.S. Pat. No. 5,726,154 show that calcitonin in anaqueous liquid composition comprising SDS (sodium dodecyl sulfate, asurfactant) and an organic acid is stable for at least 6 months.Similarly, the surfactant compositions of the present invention haveimproved stabilizing characteristics when admixed with a drug. Noorganic acid is required in these formulations. For example, thecomposition of the invention maintains the stability of proteins andpeptide therapeutics for about 6 months, or more, when maintained atabout 4° C. to 25° C.

The stability of the surfactant compositions are, in part, due to theirhigh no observable adverse effect level (NOAEL). The EnvironmentalProtection Agency (EPA) defines the no observable adverse effect level(NOAEL) as the exposure level at which there are no statistically orbiologically significant increases in the frequency or severity ofadverse effects between the exposed population and its appropriatecontrol. Hence, the term, “no observable adverse effect level” (orNOAEL) is the greatest concentration or amount of a substance, found byexperiment or observation, which causes no detectable adverse alterationof morphology, functional capacity, growth, development, or life span ofthe target organism under defined conditions.

The Food and Agriculture Organization (FAO) of the United Nations of theWorld Health Organization (WHO) has shown that some alkyl glycosideshave very high NOAELs, allowing for increased consumption of these alkylglycosides without any adverse effect. This report can be found on theworld wide web at inchem.org/documents/jecfa/jecmono/v10je11.htm. Forexample, the NOAEL for sucrose dodecanoate, a sucrose ester used in foodproducts, is about 20-30 grams/kilogram/day, e.g., a 70 kilogram person(about 154 lbs.) can consume about 1400-2100 grams (or about 3 to 4.6pounds) of sucrose dodecanoate per day without any observable adverseeffect. Typically, an acceptable daily intake for humans is about 1% ofthe NOAEL, which translates to about 14-21 grams, or 14 millionmicrograms to 21 million micrograms, per day, indefinitely. Definitionsof NOAELs and other related definitions can be found on the world wideweb at epa.gov/OCEPAterms. Thus, although some effects may be producedwith alkyl glycoside levels anticipated in the present invention, thelevels are not considered adverse, or precursors to adverse effects.

Accordingly, a subject treated with surfactant compositions of theinvention having at least one alkyl glycoside, e.g., tetradecylmaltoside(TDM; or Intravail A), at a concentration of about 0.125% by weight ofalkyl glycoside two times per day, or three times per day, or moredepending on the treatment regimen consumes about 200 to 300 microgramsper day total of TDM. So, the effective dose of the TDM is at least1000× fold lower than (i.e., 1/1000) of the NOAEL, and falls far below1% of the NOAEL, which is the acceptable daily intake; or in this caseabout 1/50,000 of the acceptable daily intake. Stated another way, alkylglycosides of the present invention have a high NOAEL, such that theamount or concentration of alkyl glycosides used in the presentinvention do not cause an adverse effect and can be safely consumedwithout any adverse effect.

The surfactant compositions of the invention are also stable becausethey are physiologically non-toxic and non-irritants. The term,“nontoxic” means that the alkyl glycoside molecule has a sufficientlylow toxicity to be suitable for human administration and consumption.Preferred alkyl glycosides are non-irritating to the tissues to whichthey are applied. Any alkyl glycoside used should be of minimal or notoxicity to the cell, such that it does not cause damage to the cell.Yet, toxicity for any given alkyl glycoside may vary with theconcentration of alkyl glycoside used. It is also beneficial if thealkyl glycoside chosen is metabolized or eliminated by the body and ifthis metabolism or elimination is done in a manner that will not beharmfully toxic. The term, “non-irritant” means that the agent does notcause inflammation following immediate, prolonged or repeated contactwith the skin surface or mucous membranes.

Moreover, one embodiment of the surfactant compositions, in particular,the sucrose esters, serve as anti-bacterial agents. An agent is an“anti-bacterial” agent or substance if the agent or its equivalentdestroy bacteria, or suppress bacterial growth or reproduction. Theanti-bacterial activity of sucrose esters and their fatty acids havebeen reported. Tetsuaki et al. (1997) “Lysis of Bacillus subtilis cellsby glycerol and sucrose esters of fatty acids,” Applied andEnvironmental Microbiology, 53(3):505-508. Watanabe et al. (2000)describe that galactose and fructose laureates are particularlyeffective carbohydrate monoesters. Watanabe et al., (2000)“Antibacterial carbohydrate monoesters suppressing cell growth ofStreptococcus mutan in the presence of sucrose,” Curr Microbiol 41(3):210-213. Hence, the present invention is not limited to the sucroseester described herein, but encompasses other carbohydrate esters,including galactose and fructose esters, that suppress bacterial growthand reproduction.

In general, all useful antimicrobial agents are toxic substances. SeeSutton and Porter (2002), “Development of the antimicrobialeffectiveness test as USP Chapter <51>,” 56(6): 300-311, which isincorporated herein by reference in its entirety. For example, commonlyused antimicrobial agents such as benzalkonium chloride are highly toxicas demonstrated by electron micrograph studies in which significantdisruption of the mucociliary surfaces are observed at concentrations ofbenzalkonium far below what is commonly used in intranasal formulations.See for example Sebahattin Cureoglu, Murat Akkus, Üstün Osma, MehmetYaldiz, Faruk Oktay, Belgin Can, Cengiz Güven, Muhammet Tekm, and FarukMeriç (2002), “The effect of benzalkonium chloride an electronmicroscopy study,” Eur Arch Otorhinolaryngol 259:362-364.

The surfactant compositions of the invention are typically present at alevel of from about 0.01% to 20% by weight. More preferred levels ofincorporation are from about 0.01% to 5% by weight, from about 0.01% to2% by weight, from about 0.01% to 1%, most preferably from about 0.01%to 0.125% by weight. The surfactant is preferably formulated to becompatible with other components present in the composition. In liquid,or gel, or capsule, or injectable, or spray compositions the surfactantis most preferably formulated such that it promotes, or at least doesnot degrade, the stability of any protein or enzyme in thesecompositions. Further, the invention optimizes the concentration bykeeping the concentration of absorption enhancer as low as possible,while still maintaining the desired effect.

The compositions of the invention when administered to the subject,yield enhanced mucosal delivery of the biologically active compound(s),or drug, with a peak concentration (or Cmax) of the compound(s) in atissue, or fluid, or in a blood plasma of the subject that is about 15%,20%, or 50% or greater as compared to a Cmax of the compound(s) in atissue (e.g., CNS), or fluid, or blood plasma following intramuscularinjection of an equivalent concentration of the compound(s) to thesubject.

The measure of how much of the drug or compound(s) reaches thebloodstream in a set period of time, e.g., 24 hours can also becalculated by plotting drug blood concentration at various times duringa 24-hour or longer period and then measuring the area under the curve(AUC) between 0 and 24 hours. Similarly, a measure of drug efficacy canalso be determined from a time to MAXIMAL CONCENTRATION (TMAX) OF THEBIOLOGICALLY ACTIVE compound(s) in a tissue (e.g., CNS) or fluid or inthe blood plasma of the subject between about 0.1 to 1.0 hours. Thetherapeutic compositions of the invention increase the speed of onset ofdrug action (i.e., reduce Tmax) by a factor of about 1.5-fold to 2-fold.

Also, the therapeutic compositions or formulations of the invention canbe administered or delivered to a subject in need systemically orlocally. Suitable routes may, for example, include oral, ocular, nasal,nasolacrimal, inhalation or pulmonary, oral cavity (sublingual or Buccalcell), transmucosal administration, vaginal, rectal, parenteraldelivery, including intramuscular, subcutaneous, intravenous,intraperitoneal, or CSF delivery. Moreover, the mode of delivery e.g.,liquid, gel, tablet, spray, etc. will also depend on the method ofdelivery to the subject.

Additionally, the therapeutic compositions of the invention can consistof a pharmaceutically acceptable carrier. A “pharmaceutically acceptablecarrier” is an aqueous or non-aqueous agent, for example alcoholic oroleaginous, or a mixture thereof, and can contain a surfactant,emollient, lubricant, stabilizer, dye, perfume, preservative, acid orbase for adjustment of pH, a solvent, emulsifier, gelling agent,moisturizer, stabilizer, wetting agent, time release agent, humectant,or other component commonly included in a particular form ofpharmaceutical composition. Pharmaceutically acceptable carriers arewell known in the art and include, for example, aqueous solutions suchas water or physiologically buffered saline or other solvents orvehicles such as glycols, glycerol, and oils such as olive oil orinjectable organic esters. A pharmaceutically acceptable carrier cancontain physiologically acceptable compounds that act, for example, tostabilize or to increase the absorption of the specific inhibitor, forexample, carbohydrates, such as glucose, sucrose or dextrans,antioxidants, such as ascorbic acid or glutathione, chelating agents,low molecular weight proteins or other stabilizers or excipients. Apharmaceutically acceptable carrier can also be selected from substancessuch as distilled water, benzyl alcohol, lactose, starches, talc,magnesium stearate, polyvinylpyrrolidone, alginic acid, colloidalsilica, titanium dioxide, and flavoring agents.

Additionally, to decrease susceptibility of a peptide drug to hydrolyticcleavage in compositions containing alkyl saccharides or saccharidealkyl esters, various oxygen atoms within the drugs can be substitutedfor by sulfur (Defaye, J. and Gelas, J. in Studies in Natural ProductChemistry (Atta-ur-Rahman, ed.) Vol. 8, pp. 315-357, Elsevier,Amsterdam, 1991). For example, the heteroatom of the sugar ring can beeither oxygen or sulfur, or the linkage between monosaccharides in anoligosaccharide can be oxygen or sulfur (Horton, D. and Wander, J. D.,“Thio Sugars and Derivatives,” The Carbohydrates: Chemistry andBiochemistry, 2d. Ed. Vol. IB, (W. Reyman and D. Horton eds.), pp.799-842, (Academic Press, New York), (1972)). Oligosaccharides can haveeither α (alpha) or β (beta) anomeric configuration (see Pacsu, E., etal. in Methods in Carbohydrate Chemistry (R. L. Whistler, et al., eds.)Vol. 2, pp. 376-385, Academic Press, New York 1963).

A composition of the invention can be prepared in tablet form by mixinga therapeutic agent or drug and one alky glycoside and/or saccharidealkyl ester according to the invention, and an appropriatepharmaceutical carrier or excipient, for example mannitol, corn starch,polyvinylpyrrolidone or the like, granulating the mixture and finallycompressing it in the presence of a pharmaceutical carrrier such as cornstarch, magnesium stearate or the like. If necessary, the formulationthus prepared may include a sugar-coating or enteric coating or coveredin such a way that the active principle is released gradually, forexample, in the appropriate pH medium.

The term “enteric coating,” is a polymer encasing, surrounding, orforming a layer, or membrane around the therapeutic composition or core.Also, the enteric coating can contain a drug which is compatible orincompatible with the coating. One tablet composition may include anenteric coating polymer with a compatible drug which dissolves orreleases the drug at higher pH levels (e.g., pH greater than 4.0,greater than 4.5, greater than 5.0 or higher) and not at low pH levels(e.g., pH 4 or less); or the reverse.

In a preferred embodiment, the dose dependent release form of theinvention is a tablet comprising: (a) a core comprising: (i) atherapeutic agent or drug; (ii) a surfactant comprising at least onealkyl glycoside and/or saccharide alkyl ester; and (b) at least onemembrane coating surrounding the core, wherein the coating is animpermeable, permeable, semi-permeable or porous coating and becomesmore permeable or porous upon contacting an aqueous environment of adefined pH. The term “membrane” is synonymous with “coating,” orequivalents thereof.

The terms are used to identify a region of a medicament, for example, atablet, that is impermeable, permeable, semi-permeable or porous to anaqueous solution(s) or bodily fluid(s), and/or to the therapeuticagent(s) or drug(s) encapsulated therein. If the membrane is permeable,semi-permeable or porous to the drug, the drug can be released throughthe openings or pores of the membrane in solution or in vivo. The porousmembrane can be manufactured mechanically (e.g., drilling microscopicholes or pores in the membrane layer using a laser), or it can beimparted due to the physiochemical properties of the coating polymer(s).Membrane or coating polymers of the invention are well known in the art,and include cellulose esters, cellulose diesters, cellulose triesters,cellulose ethers, cellulose ester-ether, cellulose acylate, cellulosediacylate, cellulose triacylate, cellulose acetate, cellulose diacetate,cellulose triacetate, cellulose acetate propionate, and celluloseacetate butyrate. Other suitable polymers are described in U.S. Pat.Nos. 3,845,770, 3,916,899, 4,008,719, 4,036,228 and 4,11210 which areincorporated herein by reference.

Further, the enteric coating according to the invention can include aplasticizer, and a sufficient amount of sodium hydroxide (NaOH) toeffect or adjust the pH of the suspension in solution or in vivo.Examples of plasticizers include triethyl citrate, triacetin, tributylsebecate, or polyethylene glycol. Other alkalizing agents, includingpotassium hydroxide, calcium carbonate, sodium carboxymethylcellulose,magnesium oxide, and magnesium hydroxide can also be used to effect oradjust the pH of the suspension in solution or in vivo.

Accordingly, in one embodiment, an enteric coating can be designed torelease a certain percentage of a drug or drugs in certain mediums witha certain pH or pH range. For example, the therapeutic composition ofthe invention may include at least one enteric coating encasing orprotecting at least one drug which is chemically unstable in an acidicenvironment (e.g., the stomach). The enteric coating protects the drugfrom the acidic environment (e.g., pH<3), while releasing the drug inlocations which are less acidic, for example, regions of the small andlarge intestine where the pH is 3, or 4, or 5, or greater. A medicamentof this nature will travel from one region of the gastrointestinal tractto the other, for example, it takes about 2 to about 4 hours for a drugto move from the stomach to the small intestine (duodenum, jejunum andileum). During this passage or transit, the pH changes from about 3(e.g., stomach) to 4, or 5, or to about a pH of 6 or 7 or greater. Thus,the enteric coating allows the core containing the drug to remainsubstantially intact, and prevents premature drug release or the acidfrom penetrating and de-stabilizing the drug.

Examples of suitable enteric polymers include but are not limited tocellulose acetate phthalate, hydroxypropylmethylcellulose phthalate,polyvinylacetate phthalate, methacrylic acid copolymer, shellac,cellulose acetate trimellitate, hydroxypropylmethylcellulose acetatesuccinate, hydroxypropylmethylcellulose phthalate, cellulose acetatephthalate, cellulose acetate succinate, cellulose acetate malate,cellulose benzoate phthalate, cellulose propionate phthalate,methylcellulose phthalate, carboxymethylethylcellulose,ethylhydroxyethylcellulose phthalate, shellac, styrene-acrylic acidcopolymer, methyl acrylate-acrylic acid copolymer, methylacrylate-methacrylic acid copolymer, butyl acrylate-styrene-acrylic acidcopolymer, methacrylic acid-methyl methacrylate copolymer, methacrylicacid-ethyl acrylate copolymer, methyl acrylate-methacrylic acid-octylacrylate copolymer, vinyl acetate-maleic acid anhydride copolymer,styrene-maleic acid anhydride copolymer, styrene-maleic acid monoestercopolymer, vinyl methyl ether-maleic acid anhydride copolymer,ethylene-maleic acid anhydride copolymer, vinyl butyl ether-maleic acidanhydride copolymer, acrylonitrile-methyl acrylate-maleic acid anhydridecopolymer, butyl acrylate-styrene-maleic acid anhydride copolymer,polyvinyl alcohol phthalate, polyvinyl acetal phthalate, polyvinylbutylate phthalate and polyvinyl acetoacetal phthalate, or combinationsthereof. One skilled in the art will appreciate that other hydrophilic,hydrophobic and enteric coating polymers may be readily employed, singlyor in any combination, as all or part of a coating according to theinvention.

The therapeutic compositions of the invention in the form of a tabletcan have a plurality of coatings, for example, a hydrophilic coating(e.g., hydroxypropylmethyl-cellulose), and/or a hydrophobic coating(e.g., alkylcelluloses), and/or an enteric coating. For example, thetablet core can be encases by a plurality of the same type of coating,or a plurality of different types of coating selected from ahydrophilic, hydrophobic or enteric coating. Hence, it is anticipatedthat a tablet can be designed having at least one, but can have morethan one layer consisting of the same or different coatings dependent onthe target tissue or purpose of the drug or drugs. For example thetablet core layer may have a first composition enclosed by a firstcoating layer (e.g., hydrophilic, hydrophobic, or enteri-coating), and asecond same or different composition or drug having the same ordifferent dosage can be enclosed in second coating layer, etc. Thislayering of various coatings provides for a first, second, third, ormore gradual or dose dependent release of the same or different drugcontaining composition.

In a preferred embodiment, a first dosage of a first composition of theinvention is contained in a tablet core and with an enteric-coating suchthat the enteric-coating protects and prevents the composition containedtherein from breaking down or being released into the stomach. Inanother example, the first loading dose of the therapeutic compositionis included in the first layer and consists of from about 10% to about40% of the total amount of the total composition included in theformulation or tablet. In a second loading dose, another percentage ofthe total dose of the composition is released. The inventioncontemplates as many time release doses as is necessary in a treatmentregimen. Thus, in certain aspects, a single coating or plurality ofcoating layers is in an amount ranging from about 2% to 6% by weight,preferably about 2% to about 5%, even more preferably from about 2% toabout 3% by weight of the coated unit dosage form.

Accordingly, the composition preparations of the invention make itpossible for contents of a hard capsule or tablet to be selectivelyreleased at a desired site the more distal parts of thegastro-intestinal tract (e.g., small and large intestine) by selectingthe a suitable pH-soluble polymer for a specific region. Mechanicalexpulsion of the composition preparations may also be achieved byinclusion of a water absorbing polymer that expands upon waterabsorption within a hard semi-permeable capsule thus expellingcomposition through an opening in the hard capsule.

Drugs particularly suited for dose dependent time release include butare not limited to insulin like growth factor-I (IGF-I),somatomedin-C(SM-C; diabetes, nerve function, renal function), insulin(diabetes), calcitonin (osteoporosis), leptin (obesity; infertility),leptin derived short peptide (OB-3), hGH (AIDs wasting, dwarfism), humanparathyroid hormone (PTH) (osteoporosis), melatonin (sleep), GLP-1 orGlucagon-like peptide-1 (diabetes), GiP (diabetes), pituitary adenylatecyclase-activating polypeptide (PACAP) and islet function (diabetes),GM-1 ganglioside, (Alzheimers), nerve growth factor (NGF), (Alzheimers),nafarelin (endometriosis), Synarel® (nafarelin acetate nasal solution),(D-tryp6)-LHRH (fertility), FGF (duodenal ulcer, macular degeneration,burns, wounds, spinal cord injuries, repair of bone and cartilagedamage), VEGF antagonists (to block the receptor), VEGF (agonist)neonatal distress syndrome; ALS), leuprolide (prostate and breastcancer), interferon-alpha (chronic hepatitis C), low molecular weightheparin (blood clotting, deep vein thrombosis), PYY (obesity), LHRHantagonists (fertility), LH (luteinizing hormone), ghrelin antagonists(obesity), KGF (Parkinson's), GDNF (Parkinsons), G-CSF (erythropoiesisin cancer), Imitrex (migraine), Integrelin (anticoagulation), Natrecor®(congestive heart failure), human B-type natriuretic peptide (hBNP),SYNAREL® (Searl; nafarelin acetate nasal solution), Sandostatin (growthhormone replacement), Forteo (osteoporosis), DDAVP® Nasal Spray(desmopressin acetate), Cetrotide® (cetrorelix acetate for injection),Antagon™ (ganirelix acetate), Angiomax (bivalirudin; thrombininhibitor), Accolate® (zafirlukast; injectable), Exendin-4 (Exanatide;diabetes), SYMLIN® (pramlintide acetate; synthetic amylin; diabetes),desmopressin, glucagon, ACTH (corticotrophin), C-peptide of insulin,GHRH and analogs (GnRHa), growth hormone releasing hormone, oxytocin,corticotropin releasing hormone (CRH), atrial natriuretic peptide (ANP),thyroxine releasing hormone (TRHrh), follicle stimulating hormone (FSH),prolactin, tobramycin ocular (corneal infections), Vasopressin,desmopresin, Fuzeon (Roche; HIV fusion inhibitor MW 4492), thymalfasin,and Eptifibatide.

Further, it will be understood by one skilled in the art, that thespecific dose level and frequency of dosage for any particular subjectin need of treatment may be varied and will depend upon a variety offactors including the activity of the specific compound employed, themetabolic stability and length of action of that compound, the age, bodyweight, general health, sex, diet, mode and time of administration, rateof excretion, drug combination, the severity of the particularcondition, and the host undergoing therapy.

It has been shown that alkyl glycosides, particularly alkylmaltosidesand more specifically, dodecylmaltoside (DDM) and tetradecylmaltoside(TDM), stabilize insulin in solution and prevent aggregation of thepeptide. Hovgaard et al., “Insulin Stabilization and GI absorption,” J.Control. Rel., 19 (1992) 458-463, cited in Hovgaard et al.,“Stabilization of insulin by alkylmaltosides: A spectroscopicevaluation,” Int. J. Pharmaceutics 132 (1996) 107-113 (hereinafter,“Hovgaard-1”). Further, Hovgaard-1 shows that even after 57 days, theDDM-insulin complex remained stable and possessed nearly full biologicalactivity. It is postulated that the stability of the complex is due tothe length of the alkyl group (number of carbon atoms) and the higherratio of DDM to insulin ratio the better (e.g., 4:1 and 16:1; see FIG. 1in Hovgaard 1). However, according to Hovgaard-1, although theDDM-insulin complex was stable, the same stability was not shown forother maltosides. Yet, in a related study, Hovgaard et al. (1996)demonstrated that when DDM-insulin was orally administered to animals invivo, bioavailability of the complex was weak (e.g., 0.5%-1%bioavailability). Hovgaard et al., “Stabilization of insulin byalkylmaltoside. B. Oral absorption in vivo in rats,” Int. J.Pharmaceutics 132 (1996) 115-121 (Hovgaard-2). Hence, an improved aspectof the invention is that the surfactant increases the bioavailability ofa drug to the target tissues, organs, system etc., as well as increasedrug stability.

Accordingly, one aspect of the invention is to provide therapeuticcompositions having at least one drug and one surfactant, wherein thesurfactant further consists of at least one alkyl glycoside and/orsaccharide alkyl ester formulation which enhances the bioavailability ofthe drug. Determining the bioavailability of drug formulations isdescribed herein. As used herein, “bioavailability” is the rate andextent to which the active substance, or moiety, which reaches thesystemic circulation as an intact drug. The bioavailability of any drugwill depend on how well is adsorbed and how much of it escapes beingremoved from the liver.

To determine absolute bioavailability, the tested drug and mode ofadministration is measured against an intravenous reference dose. Thebioavailability of the intravenous dose is 100% by definition. Forexample, animals or volunteering humans are given an intravenousinjections and corresponding oral doses of a drug. Urinary or plasmasamples are taken over a period of time and levels of the drug over thatperiod of time are determined.

The areas under the curve (AUC), of the plasma drug concentration versustime curves, are plotted for both the intravenous and the oral doses,and calculation of the bioavailability of both formulations is by simpleproportion. For example, if the same intravenous and oral doses aregiven, and the oral AUC is 50% of the intravenous AUC, thebioavailability of the oral formulation is 50%. Note that thebioavailability of any drug is due to many factors including incompleteabsorption, first pass clearance or a combination of these (discussedmore below). Further, the peak concentration (or C_(max)) of the plasmadrug concentration is also measured to the peak concentration (C_(max))of the plasma drug concentration following intramuscular (IM) injectionof an equivalent concentration the drug. Moreover, the time to maximalconcentration (or t_(max)) of the plasma drug is about 0.1 to 1.0 hours.

To determine the relative bioavailability of more than one formulationof a drug (e.g., an alkyl glycoside or saccharide alkyl ester drugformulation), bioavailability of the formulations are assessed againsteach other as one or both drugs could be subject to first pass clearance(discussed more below) and thus undetected. For example, a first oralformulation is assessed against a second oral formulation. The secondformulation is used as a reference to assess the bioavailability of thefirst. This type of study provides a measure of the relative performanceof two formulations in getting a drug absorbed.

Bioavailabilities of drugs are inconsistent and vary greatly from onedrug to the next. For example, the bioavailability of MIACALCIN® (salmoncalcitonin from Novartis) nasal spray, a prescription medication for thetreatment of postmenopausal osteoporosis in women, has a meanbioavailability of about 3% (range is 0.3%-30.6%; see FIG. 1). TheMIACALCIN® product information sheet can be found on the world wide webat miacalcin.com/info/howWorks/index.jsp anddrugs.com/PDR/Miacalcin_Nasal_Spray.html. The data on MIACALCIN®, whichwas obtained by various investigators using different methods and humansubjects, show great variability in the drug's bioavailability, e.g., innormal volunteers only ˜3% of the nasally administered dose isbioavailable, as compared to the same dose administered by intramuscularinjection (MIACALCIN® product insert). This represents two orders of amagnitude in variability and is undesirable to the consumer.

Poor bioavailability of a drug can also be observed in NASCOBAL®(Nastech), or cyanocobalamin, which is used for the treatment andmaintenance of the hematologic status of patients who are in remissionfollowing intramuscular vitamin B₁₂ therapies. The gel formulation wasadministered intranasally and the bioavailability of B₁₂ was compared tointramuscular B₁₂ injections. The peak concentrations of B₁₂ (or theTmax) was reached in 1-2 hours after intranasal administration, andrelative to the intramuscular injection, the bioavailability of B₁₂nasal gel was found to be about 8.9% (90% confidence intervals, 7.1% to11.2%).

The alkyl glycosides or sucrose esters of the present invention includeany compounds now known or later discovered. Drugs which areparticularly well suited for admixture with the alkyl glycosides and/orsaccharide alkyl esters of the invention are those that are difficult toadminister by other methods, e.g., drugs that are degraded in thegastrointestinal (GI) tract or those that are not absorbed well from theGI tract, or drugs that can be self-administered via the ocular, nasal,nasolacrimal, inhalation, or CSF delivery route instead of traditionalmethods such as injection. Some specific examples include peptides,polypeptides, proteins, nucleic acids and other macromolecules, forexample, peptide hormones, such as insulin and calcitonin, enkephalins,glucagon and hypoglycemic agents such as tolbutamide and glyburide, andagents which are poorly absorbed by enteral routes, such asgriseofulvin, an antifungal agent. Other compounds include, for example,nicotine, interferon (e.g., alpha, beta, gamma), PYY, GLP-1, syntheticexendin-4 (Exenatide), parathyroid hormone, and human growth hormone orother low molecular weight peptides and proteins.

Alternatively, bioavailability of a drug can be determined by measuringthe levels of the drug's first pass clearance by the liver. Alkylglycosides and/or saccharide alkyl ester compositions of the inventionadministered intranasally or via oral cavity (sublingual or Buccal cell)do not enter the hepatic portal blood system, thereby avoiding firstpass clearance by the liver. Avoiding first past clearance of theseformulations by the liver is described herein. The term, “first passliver clearance” is the extent to which the drug is removed by the liverduring its first passage in the portal blood through the liver to thesystemic circulation. This is also called first pass metabolism or firstpass extraction.

The two major routes of drug elimination from the body are excretion bythe kidneys whereby the drug is unchanged; and elimination by the liver,whereby the drug is metabolized. The balance between these two routesdepends on the relative efficiency of the two processes. The presentinvention describes herein elimination by the liver or liver clearance.First pass liver clearance is described by Birkett et al (1990 and1991), which is incorporated by reference in its entirety. Birkett etal., Aust Prescr, 13(1990):88-9; and Birkett et al., Austra Prescr14:14-16 (1991).

Blood carrying drug from the systemic circulation enter the liver viathe portal vein, and the liver in turn extracts a certain percentage orratio (i.e., 0.5 or 50%) of that drug. The remainder left over (i.e.,0.2 or 20%) re-enters the systemic circulation via the hepatic vein.This rate of clearance of the drug is called the hepatic extractionratio. It is the fraction of the drug in the blood which is irreversiblyremoved (or extracted) during the first pass of the blood through theliver. If no drug is extracted, the hepatic extraction ratio is zero.Conversely, if the drug is highly extracted in the first pass throughthe liver, the hepatic extraction ratio may be as high as 100% or 1.0.In general, clearance of the drug by the liver depends then on the rateof delivery of that drug to the liver (or the hepatic blood flow), andon the efficiency of removal of that drug (or the extraction ratio).

Therefore, the net equation used to determine hepatic clearance is:(hepatic clearance−blood flow)=(unbound fraction*intrinsicclearance)/blood flow+(unbound fraction*intrinsic clearance)  (1)

The “unbound fraction” of drug is dependent on how tightly the drug isbound to proteins and cells in the blood. In general, it is only thisunbound (or free) drug which is available for diffusion from the bloodinto the liver cell. In the absence of hepatic blood flow and proteinbinding, the “intrinsic clearance” is the ability of the liver to remove(or metabolize) that drug. In biochemical terms, it is a measure ofliver enzyme activity for a particular drug substrate. Again, althoughintrinsic clearance can be high, drugs cannot be cleared more rapidlythan that presented to the liver. In simple terms, there are twosituations: where liver enzyme activity is very high or very low (i.e.,high extraction ratio or low extraction ratio).

When liver enzyme activity is low, the equation simplifies to:hepatic clearance=unbound fraction*intrinsic clearance  (2)

Clearance then is independent of blood flow, but instead dependsdirectly on the degree of protein binding in the blood and the activityof drug metabolizing enzymes towards that drug.

In contrast, when liver enzyme activity is high, the equation is:hepatic clearance=liver blood flow  (3)

In this scenario, because the enzymes are so active the liver removesmost of the drug presented to it and the extraction ratio is high. Thus,the only factor determining the actual hepatic clearance is the rate ofsupply of drug to the liver (or hepatic blood flow).

First pass liver clearance is important because even small changes inthe extraction of drugs can cause large changes in bioavailability. Forexample, if the bioavailability of drug A by oral administration is 20%by the time it reaches the systemic circulation, and the same drug A byintravenous administration is 100%, absent no other complicatingfactors, the oral dose will therefore have to be 5 times the intravenousdose to achieve similar plasma concentrations.

Secondly, in some instances where liver enzyme activity is very high,drug formulations should be designed to have the drug pass directlythrough to the systemic circulation and avoid first pass liver clearanceall together. For example, drugs administered intranasally, sublingual,buccal, rectal, vagina, etc. directly enter the systemic circulation anddo not enter the hepatic portal blood circulation to be partially orfully extracted by the liver. Alternatively, where drugs cannot beadministered by the above means, a tablet with at least oneenteric-coating layer to prevent release of the drug in the stomach(i.e., highly acidic environment) is provided. Thus, an objective of theinvention is to administer drugs using these alternative routes.

Additionally, first pass liver clearance is an important factor becausemany patients are on more than one drug regimen, and this may cause druginteractions which increase or decrease liver enzyme activity; therebyincreasing or decreasing metabolism (increasing or decreasing thehepatic extraction ratio) of the drug of interest.

Hence, therapeutic compositions of the invention can be administereddirectly to the systemic circulatory system and avoid first pass liverclearance. Avoiding first pass clearance assures that more of the drugwill be available to the system. Stated another way, by avoiding firstpass liver clearance, the bioavailability of the drug is increased.

The present invention also relates to methods of increasing absorptionof a low molecular compound into the circulatory system of a subjectcomprising administering via the oral, ocular, nasal, nasolacrimal,inhalation, or the CSF delivery route the compound and an absorptionincreasing amount of a suitable nontoxic, nonionic alkyl glycosidehaving a hydrophobic alkyl joined by a linkage to a hydrophilicsaccharide.

The composition formulation is appropriately selected according to theadministration route, such as oral administration (oral preparation),external administration (e.g., ointment), injection (preparations forinjection), and mucosal administration (e.g., buccal and suppository)etc. For example, excipients (e.g., starch, lactose, crystallinecellulose, calcium lactate, magnesium aluminometasilicate and anhydroussilicate), disintegrators (e.g., carboxymethylcellulose and calciumcarboxymethylcellulose), lubricants (e.g., magnesium stearate and talc),coating agents (e.g., hydroxyethylcellulose), and flavoring agents canbe used for oral and mucosal formulations; whereas, solubilizers andauxiliary solubilizers capable of forming aqueous injections (e.g.,distilled water for injection, physiological saline and propyleneglycol), suspending agents (e.g., surfactant such as polysorbate 80), pHregulators (e.g., organic acid and metal salt thereof) and stabilizersare used for injections; and aqueous or oily solubilizers and auxiliarysolubilizers (e.g., alcohols and fatty acid esters), tackifiers (e.g.,carboxy vinyl polymer and polysaccharides) and emulsifiers (e.g.,surfactant) are used for external agents. The drug and the alkylglycoside can be admixed, mixed, or blended along with the aboveexcipients, disintegrators, coating polymers, solubilizers, suspendingagents, etc., prior to administration, or they can be administeredsequentially, in either order. It is preferred that they be mixed priorto administration.

The term, “mucosal delivery-enhancing agent” includes agents whichenhance the release or solubility (e.g., from a formulation deliveryvehicle), diffusion rate, penetration capacity and timing, uptake,residence time, stability, effective half-life, peak or sustainedconcentration levels, clearance and other desired mucosal deliverycharacteristics (e.g., as measured at the site of delivery, or at aselected target site of activity such as the bloodstream or centralnervous system) of a compound(s) (e.g., biologically active compound).Enhancement of mucosal delivery can occur by any of a variety ofmechanisms, including, for example, by increasing the diffusion,transport, persistence or stability of the compound, increasing membranefluidity, modulating the availability or action of calcium and otherions that regulate intracellular or paracellular permeation,solubilizing mucosal membrane components (e.g., lipids), changingnon-protein and protein sulfhydryl levels in mucosal tissues, increasingwater flux across the mucosal surface, modulating epithelial junctionphysiology, reducing the viscosity of mucus overlying the mucosalepithelium, reducing mucociliary clearance rates, and other mechanisms.

Exemplary mucosal delivery enhancing agents include the following agentsand any combinations thereof:

-   -   (a) an aggregation inhibitory agent;    -   (b) a charge-modifying agent;    -   (c) a pH control agent;    -   (d) a degradative enzyme inhibitory agent;    -   (e) a mucolytic or mucus clearing agent;    -   (f) a ciliostatic agent;    -   (g) a membrane penetration-enhancing agent selected from:        -   (i) a surfactant;        -   (ii) a bile salt;        -   (ii) a phospholipid additive, mixed micelle, liposome, or            carrier;        -   (iii) an alcohol;        -   (iv) an enamine;        -   (v) an NO donor compound;        -   (vi) a long-chain amphipathic molecule;        -   (vii) a small hydrophobic penetration enhancer;        -   (viii) sodium or a salicylic acid derivative;        -   (ix) a glycerol ester of acetoacetic acid;        -   (x) a cyclodextrin or beta-cyclodextrin derivative;        -   (xi) a medium-chain fatty acid;        -   (xii) a chelating agent;        -   (xiii) an amino acid or salt thereof;        -   (xiv) an N-acetylamino acid or salt thereof        -   (xv) an enzyme degradative to a selected membrane component;        -   (ix) an inhibitor of fatty acid synthesis;        -   (x) an inhibitor of cholesterol synthesis; and        -   (xi) any combination of the membrane penetration enhancing            agents recited in (i)-(x);    -   (h) a modulatory agent of epithelial junction physiology;    -   (i) a vasodilator agent;    -   (j) a selective transport-enhancing agent; and    -   (k) a stabilizing delivery vehicle, carrier, mucoadhesive,        support or complex-forming species with which the compound is        effectively combined, associated, contained, encapsulated or        bound resulting in stabilization of the compound for enhanced        nasal mucosal delivery, wherein the formulation of the compound        with the intranasal delivery-enhancing agents provides for        increased bioavailability of the compound in a blood plasma of a        subject.

Additional mucosal delivery-enhancing agents include, for example,citric acid, sodium citrate, propylene glycol, glycerin, ascorbic acid(e.g., L-ascorbic acid), sodium metabisulfite,ethylenediaminetetraacetic acid (EDTA) disodium, benzalkonium chloride,sodium hydroxide, and mixtures thereof. For example, EDTA or its salts(e.g., sodium or potassium) are employed in amounts ranging from about0.01% to 2% by weight of the composition containing alkyl saccharidepreservative.

Therapeutic agents or drugs of the present invention can be peptides orproteins, medically or diagnostically useful, of small to medium size,e.g., up to about 15 kD, 30 kD, 50 kD, 75 kD, etc., or a protein havingbetween about 1-300 amino acids or more. The methods of the inventionalso anticipate the use of small molecules, for example, an organiccompound that has a molecular weight of less than 3 kD, or less than 1.5kD.

The mechanisms of improved drug absorption according to the inventionare generally applicable and should apply to all such peptides orprotein, although the degree to which their absorption is improved mayvary according to the molecular weight (MW) and the physico-chemicalproperties of the peptide or protein, and the particular enhancer used.Examples of peptides or protein include vasopressin, vasopressinpolypeptide analogs, desmopressin, glucagon, corticotropin (ACTH),gonadotropin, calcitonin, C-peptide of insulin, parathyroid hormone(PTH), growth hormone (HG), human growth hormone (hGH), growth hormonereleasing hormone (GHRH), oxytocin, corticotropin releasing hormone(CRH), somatostatin or somatostatin polypeptide analogs, gonadotropinagonist or gonadotrophin agonist polypeptide analogs, human atrialnatriuretic peptide (ANP), human thyroxine releasing hormone (TRH),follicle stimulating hormone (FSH), and prolactin.

One preferred composition of the invention is the peptide drug isExenatide (or exendin-4) and an alkyl glycoside. Exenatide is asynthetic version of exendin-4, and has been used in clinical trials byAmylin™ Pharmaceuticals. Exendin-4 is a low molecular weight peptidethat is the first of a new class of therapeutic medications known asincretin mimetic agents or hormones. Incretin hormones are any ofvarious gastrointestinal (GI) hormones and factors that act as potentstimulators of insulin secretion, e.g., as gastric inhibitorypolypeptide (GIP), glucagon-like peptide-1 (GLP-1), or Exenatide, orexendin-4, or equivalents thereof.

Exendin-4 is a naturally occurring 39-amino acid peptide isolated fromsalivary secretions of the Gila Monster Lizard. Eng et al., “Isolationand characterization of exendin-4, an exendin-3 analogue, from Helodermasuspectum venom. Further evidence for an exendin receptor on dispersedacini from guinea pig pancreas,” J. Biol. Chem. 267(15):7402-7405(1992). Exenatide exhibits similar glucose lowering actions to glucagonslike peptide, or GLP-1. Exenatide is being investigated for itspotential to address important unmet medical needs of many people withtype 2 diabetes. Clinical trials suggest that Exenatide treatmentdecreases blood glucose toward target levels and is associated withweight loss. The effects on glucose control observed with Exenatidetreatment are likely due to several actions that are similar to those ofthe naturally occurring incretin hormone GLP-1 (see Example 7). Theseactions include stimulating the body's ability to produce insulin inresponse to elevated levels of blood glucose, inhibiting the release ofglucagon following meals and slowing the rate at which nutrients areabsorbed into the bloodstream. In animal studies Exenatideadministration resulted in preservation and formation of new beta cells,the insulin-producing cells in the pancreas, which fail as type 2diabetes progresses.

Use of Exenatide, incretin mimetic agents or equivalents thereof can beused to treat various forms of diabetes including but not limited tobrittle diabetes, chemical diabetes or impaired glucose tolerance,gestational diabetes, diabetes insipidus, diabetes insipidus central,diabetes insipidus nephrogenic, diabetes insipidus pituitary, latentdiabetes, lipatrophic diabetes, maturity-onset diabetes of youth (MODY),diabetes mellitus (DM), diabetes mellitus adult-onset (type 2 DM),diabetes mellitus insulin-dependent (IDDM, or type 1 DM), diabetesmellitus non-insulin dependent (NIDDM), diabetes mellitus juvenile orjuvenile-onset, diabetes mellitus ketosis-prone, diabetes mellitusketosis-resistant, diabetes mellitus malnutrition-related (MRDM),diabetes mellitus tropical or tropical pancreatic, diabetes mellitus,preclinical diabetes, or diabetes induced by various drugs e.g.,thiazide diabetes, steroid diabetes, or various diabetes animal modelincluding but not limited to alloxan diabetes and puncture diabetes.

In another aspect, therapeutic compositions of the invention are used totreat obesity. Obesity is a common problem in both adults andadolescents. For example, PYY3-36 (or AC162352) is a hormone that playsa critical role in decreasing appetites. The gut hormone fragmentpeptide PYY3-36 (PYY) reduces appetite and food intake when infused intosubjects of normal weight. Similar to the adipocyte hormone, leptin, PYYreduces food intake by modulating appetite circuits in the hypothalamus.However, in obese patients there is a resistance to the action ofleptin, thereby limiting leptin's therapeutic effectiveness. Still otherstudies show that PYY reduces food intake. Injection of PYY revealedthat they eat on average 30% less than usual, resulting in weight loss.Hence, PYY 3-36 has potential as a treatment for obesity. Amylin™Pharmaceuticals submitted an Investigational New Drug application forPYY 3-36 in 2003.

Compounds whose absorption can be increased by the method of thisinvention include any compounds now known or later discovered, inparticular drugs, or therapeutic compounds, molecules or agents that aredifficult to administer by other methods, for example, drugs that aredegraded in the gastrointestinal (GI) tract or that are not absorbedwell from the GI tract, or drugs that subjects could administer tothemselves more readily via the ocular, nasal, nasolacrimal, inhalationor pulmonary, oral cavity (sublingual or Buccal cell), or CSF deliveryroute than by traditional self-administration methods such as injection.Some specific examples include peptides, polypeptides, proteins andother macromolecules, for example, peptide hormones, such as insulin andinsulin analogs or derivatives such as Humalog™ and Novalog™, amongothers and calcitonin, enkephalins, glucagon and hypoglycemic agentssuch as tolbutamide and glyburide, and agents which are poorly absorbedby enteral routes, such as griseofulvin, an antifungal agent. Othercompounds include, for example, nicotine, interferon (e.g., alpha, beta,gamma), PYY, GLP-1, synthetic exendin-4 (Exenatide), parathyroid hormone(PTH), and human growth hormone or other low molecular weight peptidesand proteins.

As discussed herein, varying amounts of drug may be absorbed as a drugpasses through the buccal, sublingual, oropharyngeal and oesophagealpregastric portions of the alimentary canal. However, the bulk of thedrug passes into the stomach and is absorbed in the usual mode in whichenteric dosage forms such as tablets, capsules, or liquids are absorbed.As drug is absorbed from the intestines, the drug is brought directlyinto the liver, where, depending upon its specific chemical structure,it may be metabolized and eliminated by enzymes that perform the normaldetoxifying processes in liver cells. This elimination is referred to as“first-pass” metabolism or the “first-pass” effect in the liver aspreviously discussed. The resulting metabolites, most oftensubstantially or completely inactive compared to the original drug, areoften found circulating in the blood stream and subsequently eliminatedin the urine and/or feces.

Aspects of the present invention are based on the discovery thataddition of certain alkyl saccharides, when included in fast-dispersingdosage forms, modulate the proportion of drug that is subject to thefirst-pass effect, thus allowing a fixed amount of drug to exert greaterclinical benefit, or allowing a smaller amount of drug to achievesimilar clinical benefit compared to an otherwise larger dose.

Additional aspects of the invention are based on the discovery thatincreasing or decreasing the amount of specific alkyl saccharidesincluded in fast-dispersing dosage forms alters or modulates the site ofabsorption of a drug, increasing or decreasing, respectively, thatproportion of a drug that is absorbed through buccal tissue compared toother portions of the alimentary canal. In cases where it is desirableto speed the onset of drug action but preserve the normally longer Tmaxassociated with the standard oral tablet, the alkylsaccharide contentcan be reduced to attenuate buccal absorption so that a portion of thedrug is immediately absorbed buccally for rapid onset, but the rest isabsorbed through the slower gastric absorption process. In this way ithas been found that by selecting an alkylsaccharide concentration lessthan, for example 20% less than, the concentration of alkylsaccharidethat has been found by experiment to produce maximal or near maximalbuccal absorption, a broader absorption peak in the “systemic druglevel” vs time graph, overall, may be achieved where this is judged tobe clinically desirable.

As further discussed in the Examples below, addition of certainalkylsaccharides having specific alkyl chain lengths to thefast-dispersing tablets alters the pharmacokinetics of pre-gastric drugabsorption in beneficial ways. Specifically, incorporation of frombetween about 0.2%-0.3%, 0.3%-0.4%, 0.4%-0.5%, 0.5%-1.0%, 1.0%-2.0%,2.0%-3.0%, 3.0%-4.0%, 4.0%-5.0%, 5.0%-6.0%, 6.0%-7.0%, 7.0%-8.0%,9.0%-10.0% and greater than 10% of alkylglycoside alters thepharmacokinetics of pre-gastric drug absorption in beneficial ways. Inexemplary embodiments, the alkylsaccharide is dodecyl maltoside,tetradecyl maltoside and/or sucrose dodecanoate, which when incorporatedinto a fast-dispersing tablet format increases the drug that enters intosystemic circulation and decreases the drug that is eliminated by the“first-pass” effect in the liver. Additionally, the time to maximum druglevels is dramatically reduced, typically from one to six hours, toapproximately 15 to 45 minutes. For use in treating combative patientsundergoing psychotic episodes, this more rapid absorption of drug,resulting in more rapid onset of action, may be of great benefit.

Further, other aspects of the invention, are based on the discovery thatwhen certain types of fast-dissolve or fast-dispersing tablets areplaced between the cheek and gum or into close association with buccaltissue inside the mouth, an even larger proportion of drug is directlyabsorbed into systemic circulation and a smaller amount subsequentlyundergoes first pass elimination in the liver. Lastly, it has beendiscovered that a particularly favorable location within the mouth forthis effect is inside the central portion of the upper lip, between theinside of the lip and gums, directly below the nose. In exemplaryaspects, these types of fast-dissolve dosage formulations are preparedby lyophilization or vacuum drying. In an exemplary aspect, the dosageformulation is prepared in a manner that results in a dosage formulationthat is substantially porous.

The term “fast-dispersing dosage form” is intended to encompass all thetypes of dosage forms capable of dissolving, entirely or in part, withinthe mouth. However, in exemplary aspects, the fast-dispersing dosageform is a solid, fast-dispersing network of the active ingredient and awater-soluble or water-dispersible carrier matrix which is inert towardsthe active ingredient and excipients. In various embodiments, thenetwork may be obtained by lyophilizing or subliming solvent from acomposition in the solid state, which composition comprises the activeingredient, an alkyl saccharide, and a solution of the carrier in asolvent. While a variety of solvents are known in the art as beingsuitable for this use, one solvent particularly well suited for use withthe present invention is water. Water-alcohol mixtures may also beemployed where drug solubility in the mixed solvent is enhanced. Forpoorly water soluble drugs, dispersions of small drug particles can besuspended in an aqueous gel that maintains uniform distribution of thesubstantially insoluble drug during the lyophilization or sublimingprocess.

In one embodiment, the aqueous gel may be the self-assembling hydrogelsdescribed in U.S. Patent Application No. 60/957,960, formed usingselected alkylsaccharides such as sucrose mono- and di-stearate and/ortetradecyl-maltoside, incorporated herein by reference. In variousaspects, the fast-dissolve compositions of the invention disintegrateswithin 20 seconds, preferebly less than 10 seconds, of being placed inthe oral cavity.

Matrix forming agents suitable for use in fast-dissolve formulations ofthe present invention are describe throughout this application. Suchagents include materials derived from animal or vegetable proteins, suchas the gelatins, collagens, dextrins and soy, wheat and psyllium seedproteins; gums such as acacia, guar, agar, and xanthan; polysaccharides;alginates; carrageenans; dextrans; carboxymethylcelluloses; pectins;synthetic polymers such as polyvinylpyrrolidone; and polypeptide/proteinor polysaccharide complexes such as gelatin-acacia complexes. Inexemplary aspects, gelatin, particularly fish gelatin or porcine gelatinis used.

While it is envisioned that virtually any drug may be incorporated intoa fast-dissolve dosage formulation as described herein, particularlywell suited drugs include melatonin, raloxifene, olanzapene anddiphenhydramine.

Further, the therapeutic compositions of the invention also contemplatenon-peptide drugs or therapeutic agents. For example, in U.S. Pat. No.5,552,534, non-peptide compounds are disclosed which mimic or inhibitthe chemical and/or biological activity of a variety of peptides. Suchcompounds can be produced by appending to certain core species, such asthe tetrahydropyranyl ring, chemical functional groups which cause thecompounds to be at least partially cross-reactive with the peptide. Aswill be recognized, compounds which mimic or inhibit peptides are tovarying degrees cross-reactivity therewith. Other techniques forpreparing peptidomimetics are disclosed in U.S. Pat. Nos. 5,550,251 and5,288,707. The above U.S. patents are incorporated by reference in theirentirety.

The method of the invention can also include the administration, alongwith the alkyl glycoside and a protein or peptide, a protease orpeptidase inhibitor, such as aprotinin, bestatin, alpha₁ proteinaseinhibitor, soybean trypsin inhibitor, recombinant secretory leucocyteprotease inhibitor, captopril and other angiotensin converting enzyme(ACE) inhibitors and thiorphan, to aid the protein or peptide inreaching its site of activity in the body in an active state (i.e., withdegradation minimal enough that the protein is still able to functionproperly). The protease or peptidase inhibitor can be mixed with thealkyl glycoside and drug and then administered, or it can beadministered separately, either prior to or after administration of theglycoside or drug.

The invention also provides a method of lowering blood glucose level ina subject comprising administering a blood glucose-reducing amount of acomposition comprising insulin and an absorption increasing amount of asuitable nontoxic, nonionic alkyl glycoside having a hydrophobic alkylgroup joined by a linkage to a hydrophilic saccharide, therebyincreasing the absorption of insulin and lowering the level of bloodglucose. A “blood glucose-reducing amount” of such a composition is thatamount capable of producing the effect of reducing blood glucose levels,as taught herein. Preferred is an amount that decreases blood glucose tonormoglycemic or near normoglycemic range. Also preferred is an amountthat causes a sustained reduction in blood glucose levels. Even morepreferred is an amount sufficient to treat diabetes, including diabetesmellitus (DM) by lowering blood glucose level. Thus, the instant methodcan be used to treat diabetes mellitus. Preferred alkyl glycosides arethe same as those described above and exemplified in the Examples.

Also provided is a method of raising blood glucose level in a subject byadministering a blood glucose-raising amount comprising glucagons and atleast one alkyl glycoside and/or saccharide alkyl ester. When thecomposition includes insulin, it can be used to cause the known effectof insulin in the bloodstream, i.e., lower the blood glucose levels in asubject. Such administration can be used to treat diabetes mellitus, orrelated diseases. A “blood glucose-raising amount” of glucagon in such acomposition is that amount capable of producing the effect of raisingblood glucose levels. A preferred amount is that which increases bloodglucose to normoglycemic or near-normoglycemic range. Another preferableamount is that which causes a sustained rising of blood glucose levels.Even more preferred, is that amount which is sufficient to treathypoglycemia by raising blood glucose level. Thus, this method can beused to treat hypoglycemia. Preferred alkyl glycosides are the same asthose described above and exemplified in the Examples.

Similarly, when this composition includes glucagon, it can be used tocause the known effect of glucagon in the bloodstream, i.e., to raisethe blood glucose levels in a subject. Such administration can thereforebe used to treat hypoglycemia, including hypoglycemic crisis.

The invention also provides methods for ameliorating neurologicaldisorders which comprises administering a therapeutic agent to thecerebral spinal fluid (CSF). The term “neurological disorder” denotesany disorder which is present in the brain, spinal column, and relatedtissues, such as the meninges, which are responsive to an appropriatetherapeutic agent. The surprising ability of therapeutic agents of thepresent invention to ameliorate the neurological disorder is due to thepresentation of the therapeutic agent to persist in thecerebro-ventricular space. The ability of the method of the invention toallow the therapeutic agent to persist in the region of the neurologicaldisorder provides a particularly effective means for treating thosedisorders.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular subject in need of treatment maybe varied and will depend upon a variety of factors including theactivity of the specific compound employed, the metabolic stability andlength of action of that compound, the age, body weight, general health,sex, diet, mode and time of administration, rate of excretion, drugcombination, the severity of the particular condition, and the hostundergoing therapy. Generally, however, dosage will approximate thatwhich is typical for known methods of administration of the specificcompound. For example, for intranasal administration of insulin, anapproximate dosage would be about 0.5 unit/kg regular porcine insulin(Moses et al.). Dosage for compounds affecting blood glucose levelsoptimally would be that required to achieve proper glucose levels, forexample, to a normal range of about 5-6.7 mM. Additionally, anappropriate amount may be determined by one of ordinary skill in the artusing only routine testing given the teachings herein (see Examples).

Furthermore, the compositions of the invention can be administered in aformat selected from the group consisting of a drop, a spray, an aerosoland a sustained release format. The spray and the aerosol can beachieved through use of the appropriate dispenser. The sustained releaseformat can be an ocular insert, erodible microparticulates, swellingmucoadhesive particulates, pH sensitive microparticulates,nanoparticles/latex systems, ion-exchange resins and other polymericgels and implants (Ocusert, Alza Corp., California; Joshi, A., S. Pingand K. J. Himmelstein, Patent Application WO 91/19481). These systemsmaintain prolonged drug contact with the absorptive surface preventingwashout and nonproductive drug loss.

The present invention is more particularly described in the followingexamples which are intended as illustrative only since numerousmodifications and variations therein will be apparent to those skilledin the art. The following examples are intended to illustrate but notlimit the invention.

Example 1 Alkyl Glycoside and/or Sucrose Ester Formulations do not CauseMucosa Irritation or Disruption

The nasal mucosa is highly vascularized and hence optimal for high drugpermeation. Moreover, absorption of drug(s) through the nasal mucosa isavailable to the central nervous system (CNS). Although localapplication of drugs is desirable, a challenge for this method ofadministration is mucosal irritancy.

A formulation consisting of an alkyl glycoside (0.125% TDM) in acommercial over-the-counter (OTC) nasal saline was administered in vivoto human nasal epithelium over a period of over one month. The 0.125%TDM formulation is compared to the control, namely the same commercial(OTC) nasal saline, over the same period of time. Results show thatduring and after 33 days of daily TDM administration (i.e., the durationof the study), there is no observable irritation of the nasal mucosa(data not shown). Thus, compositions of the invention are non-toxic andnon-irritable providing repeated and long-term intranasaladministration, which is beneficial for those patients with chronic andongoing disease(s).

A similar test was performed using sucrose dodecanoate, a sucrose ester.Sucrose dodecanoate is administered in vivo to human nasal epitheliumand during and after 47 days (i.e., the duration of the study), noobservable irritation was detected (data not shown). Thus, these resultsshow that alkyl glycosides and sucrose esters of the invention arenon-toxic and do not cause mucosa irritation when administered dailyover a long period of time.

Example 2 Alkyl Glycoside and/or Sucrose Ester Compositions StabilizeDrugs by Increasing Drug Bioavailability and Reducing DrugBioavailability Variance

Stability of the alkyl glycoside depends, in part, on the number ofcarbon atoms or length of the alkyl chain and other long alkyl chains,with tetradecylmaltoside (TDM) having the greatest effect; but otherhighly branched alkyl chains including DDM also have stabilizingeffects. In contrast to Hovgaard-1, which described the preference for ahigh alkyl glycoside to drug ratio, the instant invention shows thatthis ratio is much lower. For example, alkyl glycosides in the range ofabout 0.01% to about 6% by weight result in good stabilization of thedrug; whereas Hovgaard-1 shows stabilization is only achieved at muchhigher ratios of alkyl glycosides to drug (10:1 and 16:1). Even moreinteresting, alkyl glycosides of the invention in the range of about0.01% to about 6% have increased bioavailability (see FIG. 1). This isin sharp contrast to Hovgaard-2, which showed relatively lowbioavailability (0.5-1%) at the high alkyl glycoside ratios (10:1 and16:1).

FIG. 1 is a graph comparing the bioavailability of the drug MIACALCIN®(salmon calcitonin from Novartis) with and without alkyl glycoside(TDM). MIACALCIN® is a nasal spray and administered directly onto thenasal epithelium or nasal mucosa. FIG. 1 shows that MIACALCIN® minusalkyl glycoside has very low bioavailability levels in humans(MIACALCIN® product specification insert), as compared to the MIACALCIN®with alkyl glycoside as administered to rats. More specifically,intranasal delivery of MIACALCIN® with 0.125% and 0.250% alkyl glycoside(TDM) resulted in about 43% to about 90% bioavailability, respectively.The bioavailability of intranasal administration of MIACALCIN® withoutalkyl glycoside is only about 3% in humans, and was undetectable inrats, suggesting that the rat is a stringent model for estimatingintranasal drug absorption in humans. Thus, the alkyl glycoside of theinvention enhances absorption and increases bioavailability of the drug.

Furthermore, besides increasing the bioavailability of the drug, thealkyl glycoside compositions of the invention effectively decrease thebioavailability variance of the drug. FIG. 1 shows that administrationof MIACALCIN® with alkyl glycoside (0.125% or 0.25%) intranasally has abioavailability variance of +/−8%, whereas the bioavailability variancewithout alkyl glycoside is 0.3% to 30%, or a two orders of magnitudechange. The increase in bioavailability and the decrease in thebioavailability variance ensures patient-to-patient variability is alsoreduced. The results as shown in FIG. 1 are administered intranasally,however, similar results are expected for oral, buccal, vaginal, rectal,etc. delivery and at different alkyl glycoside concentrations.

Thus, contrary to the art, the alkyl glycoside compositions of theinvention, in the range of about 0.01% to about 6% result in increasedbioavailability and reduced bioavailability variance. This has nototherwise been reported.

Example 3 Ocular Administration of Alkyl Saccharides Plus InsulinProduces Hypoglycemic Effects In Vivo

Normal rats were anesthetized with a mixture of xylazine/ketamine toelevate their blood glucose levels. The elevated levels of D-glucosethat occur in response to anesthesia provide an optimal system tomeasure the systemic hypoglycemic action of drug administration, e.g.,insulin-containing eye drops. This animal model mimics the hyperglycemicstate seen in diabetic animals and humans. In the experimental animalgroup, anesthetized rats are given eye drops containing insulin. Bloodglucose levels from the experimental group are compared to anesthetizedanimals which received eye drops without insulin. The change in bloodglucose levels and the differential systemic responses reflects theeffect of insulin absorbed via the route of administration, e.g., ocularroute.

Adult male Sprague-Dawley rats (250-350 g) were fed ad libitum, andexperiments were conducted between 10:00 am. and 3:00 p.m. Rats wereanesthetized with a mixture of xylazine (7.5 mg/kg) and ketamine (50mg/kg) given intraperitoneally (IP) and allowed to stabilize for 50-90min before the administration of eye drops. Anesthesia of a normal ratwith xylazine/ketamine produces an elevation in blood glucose valueswhich provides an optimal state to determine the systemic hypoglycemicaction of insulin-containing eye drops. Blood D-glucose values weremeasured by collecting a drop of blood from the tail vein at 5-10 minintervals throughout the experiment and applying the blood to glucometerstrips (Chemstrip bG) according to directions provided with theinstrument (Accu-Chek II, Boehringer Mannheim Diagnostics; Indianapolis,Ind.). Blood D-glucose values ranged from 200 to 400 mg/dl inanesthetized nondiabetic rats.

At time 0, after a 50-90 min stabilization period, rats were given 20 μlof eye drops composed of phosphate-buffered saline (PBS) with or without0.2% regular porcine insulin and 0.125%-0.5% of the absorption enhancingalkyl glycoside (e.g., TDM) to be tested. Eye drops were instilled attime 0 using a plastic disposable pipette tip with the eyes held open,and the rat was kept in a horizontal position on a warming pad (37° C.)throughout the protocol. The rats were given additional anesthesia ifthey showed signs of awakening. Rats received in each eye 20 μl of0.125-0.5% absorption enhancer in phosphate buffered saline, pH 7.4 with(experimental) or without (control) 0.2% (50 U/ml) regular porcineinsulin (Squibb-Novo, Inc.) for a total of 2 U per animal.Octyl-β-D-maltoside, decyl-β-D-maltoside, dodecyl-D-maltoside,tridecyl-β-D-maltoside and tetradecyl-β-D-maltoside were obtained fromAnatrace, Inc. (Maumee, Ohio). Hexylglucopyranoside,heptylglucopyranoside, nonylglucopyranoside, decylsucrose anddodecylsucrose were obtained from Calbiochem, Inc. (San Diego, Calif.);Saponin, BL-9 and Brij 78 were obtained from Sigma Chemical Co. (St.Louis, Mo.).

The D-glucose levels in the blood remained elevated when the animalsreceived eye drops containing: 1) saline only; 2) 0.2% regular porcineinsulin in saline only; or 3) absorption enhancer only. However, whenrats received eye drops containing 0.2% regular porcine insulin andseveral alkylmaltoside or alkylsucrose compounds, a pronounced decreasein blood D-glucose values occurred and was maintained for up to twohours. Insulin administered ocularly with 0.5% dodecyl-β-D-maltoside(see Table I) or 0.5% decyl-β-D-maltoside (see Table III) results in aprompt and sustained fall in blood glucose levels which are maintainedin the normoglycemic (80-120 mg/dl) or near-normoglycemic (120-160mg/dl) range for the two hour duration of the experiment. Hence, atleast two alkylmaltosides are effective in achieving sufficientabsorption of insulin delivered via the ocular route to produce a promptand sustained fall in blood glucose levels in experimentallyhyperglycemic animals. The surfactant compositions of the invention aretherefore useful to achieve systemic absorption of insulin and otherpeptides/proteins, e.g., glucagon and macromolecular drugs and heparindelivered via the ocular route in the form of eye drops.

Several other alkylmaltosides are also effective as absorption enhancersfor ocular administration of insulin including 0.5% tridecylmaltoside(see Table III) and 0.125% (Table II) and 0.5% tetradecyl maltoside.These studies show that alkylmaltosides with the longer alkyl chains (ornumber of carbon atoms), e.g., dodecyl-, tridecyl- andtetradecyl-β-D-maltosides, are more effective. The increase in thenumber of carbon atoms also contributes to the greaterhydrophobic/hydrophilic structural balance and absorption enhancingeffect. The shorter alkyl chains (fewer carbon atoms) e.g.,decylmaltoside, or no, e.g., octylmaltoside, produce less absorptionenhancing activity. It is noted that the most effective alkylmaltosidesproduce effects comparable to or greater than those seen with otherabsorption enhancers such as saponin, and with the added advantage thatthey can be metabolized to nontoxic products following systemicabsorption.

The effects of the alkylmaltosides as absorption enhancers aredose-dependent, as can be seen by examining the effects of differentconcentrations ranging from 0.125-0.5% in producing a hypoglycemiceffect when combined with insulin. Whereas, 0.5% and 0.375%dodecylmaltoside appear equally effective in achieving systemicabsorption of insulin and reduction of blood glucose levels, 0.25% has asmaller and more transient effect and 0.125% is ineffective (Table I).Similarly, tridecylmaltoside also shows a dose-dependent effect inlowering blood glucose concentrations when combined with insulin, butthe effect achieved with even 0.25% of the absorption enhance issustained for the two hour time course of the experiment. Thus,dose-dependent effects of the alkylmaltosides suggest that they achieveenhancement of protein absorption via the ocular route in a gradedfashion proportional to the concentration of the agent.

TABLE I Effect of Eye Drops Containing Insulin Plus VariousConcentrations of Dodecyl Maltoside on Blood Glucose Values (in mg/dl)in Rat Dodecyl Maltoside Concentration 0.125% 0.25% 0.375% 0.50% Time(min) Blood Glucose Concentrations (mg/dl) −20 305 ± 60 271 ± 38 305 ±51 375 ± 9  −10 333 ± 58 295 ± 32 308 ± 27 366 ± 12 0 338 ± 67 323 ± 62309 ± 32 379 ± 4  30 349 ± 64 250 ± 48 212 ± 18 297 ± 18 60 318 ± 38 168± 22 134 ± 4  188 ± 25 90 325 ± 57 188 ± 55 125 ± 12 141 ± 13 120 342 ±78 206 ± 63 119 ± 19 123 ± 5 

The absorption enhancing effects of the alkyl saccharides were notconfined to the alkylmaltosides alone since dodecylsucrose (0.125%,0.25%, 0.375%) also shows a dose-dependent effect in producing ocularabsorption of insulin and reduction in blood glucose levels. This effectis observed even at 0.125% alkyl saccharide (from 335 mg/dl.+−0.26 mg/dlat time 0 min. to 150 mg/dl+−0.44 mg/dl at time 120 min.). 0.5%decylsucrose was also effective in reducing blood glucose levels, but asshown for the alkylmaltosides, a reduction in the length of the alkylchain, and hence the hydrophobic properties of the molecule, appears toreduce the potency of the alkylsucrose compounds. However, a significantand sustained reduction in blood glucose levels is achieved with 0.5%decylsucrose (from 313 mg/dl.+−0.15 mg/dl at time 0 min. to 164mg/dl+−0.51 mg/dl at time 120 min.). The absorption enhancing abilitiesof alkyl saccharides with two distinct disaccharide moieties suggeststhat it is the physicochemical properties of the compounds which arecrucial to their activity and that other alkyl saccharides, e.g.,dodecyllactose, have the right balance of properties to be equally ormore effective as absorption enhancers while retaining the metabolic andnontoxic properties of the alkylsaccharide enhancing agents. These alkylsaccharides are anticipated by the invention.

Studies with alkylglucosides were also conducted; 0.5% hexylglucosideand 0.5% heptylglucoside were ineffective at promoting insulinabsorption from the eye, but 0.5% nonylglucoside effectively stimulatedinsulin absorption and reduced blood glucose levels (from 297 mg/dl to150 mg/dl). This result once further supports that the alkyl chainlength, as well as the carbohydrate moiety, play critical roles ineffectively enhancing insulin absorption.

It should be noted that no damaging effects (i.e., non-irritants) to theocular surface were observed with any of the alkylmaltoside oralkylsucrose agents employed in these studies. Furthermore, the promptand sustained hypoglycemic effects produced by these agents incombination with insulin suggest that these absorption enhancers do notadversely affect the biological activity of the hormone, in keeping withtheir nondenaturing, mild surfactant properties.

Thus, therapeutic compositions on the invention consisting of at leastan alkyl glycoside and a drug are stable and the alkyl glycosidesenhance the absorption of the drug.

Example 4 Ocular and Intranasal Administration of TDM Plus GlucagonProduces Hypoglycemic Effects In Vivo

Since previous Examples showed that administration via eye drops of anabsorption enhancer with drug e.g., insulin results in significantabsorption of the drug via the nasolacrimal drainage system,therapeutically effective administration of insulin withalkylmaltosides, alkylsucrose and like agents by intranasaladministration is tested herein.

Tetradecylmaltoside (TDM) in combination with insulin also produced adrop in blood D-glucose levels when administered in the form of a dropintranasally as well as via a drop by the ocular route. Eye dropscontaining 0.2% regular porcine insulin with 0.125% tetradecylmaltosideare administered to rats as previously described. The administration ofthe composition produces a prompt and prominent drop in blood glucoselevels. The drop in blood glucose levels decrease even more byadministration of a nose drop containing the same concentration ofinsulin with 0.5% tetradecylmaltoside (Table II). Thus, intranasaldelivery and administration of the alkyl saccharide with drug results inlowering of blood glucose levels.

TABLE II Effect of Insulin Eye Drops, Containing 0.125% TetradecylMaltoside and Nose Drops Containing 0.5% Tetradecyl Maltoside on BloodGlucose Values in Rats Time (min) Blood Glucose (mg/dl) −20 319 −10 311Eye drops added 0 322 15 335 30 276 45 221 60 212 75 167 90 174 105 167120 208 Nose Drops Added 135 129 150 74 165 76 180 68

Example 5 Ocular Administration of Alkyl Saccharides Plus InsulinProduces Hyperglycemic Effects In Vivo

Previous studies demonstrated that insulin absorption from the eye isstimulated by saponin, BL-9 and Brij-78. BL-9 and Brij-78 areineffective at stimulating the absorption of glucagon from the eye,whereas saponin is effective. Glucagon absorption from the eye wasmeasured in rats given eye drops containing various surfactants plusglucagon (30 μg) (Eli Lilly, Indianapolis, Ind.) by monitoring anelevation in blood D-glucose levels. In these experiments, rats wereanesthetized with sodium pentobarbital rather than xylazine/ketamin.This modification of the procedure resulted in basal blood glucoselevels in the normoglycemic range and made it possible to readilymonitor the hyperglycemic action of any glucagon absorbed from the eye.

Paired animals that receive eye drops containing the surfactant alone,or glucagon alone, were compared to animals receiving eye drops with thesurfactant plus glucagon. When eyedrops containing 0.5% saponin plusglucagon are administered to rats, the level of D-glucose in blood risessignificantly, but no such effect is observed with eye drops containing0.5% BL-9 or 0.5% Brij-78 plus glucagon. Interestingly, when eye dropscontaining dodecylsucrose, decylmaltose or tridecylmaltose plus glucagonare administered to rats which were previously treated with eye dropscontaining these surfactant agents plus insulin, the glucagon isabsorbed and blood D-glucose values increase significantly (Table III).This result confirms that ocular administration of certainalkylsaccharides can enhance the absorption of drugs, including glucagonand insulin. Moreover, it is now possible to treat for a hypoglycemiccrisis using a formulation with at least an alkyl saccharide of theinvention.

TABLE III Effect of Eye Drops Containing Insulin or Glucagon and 0.5%Decyl Maltoside, 0.5% Dodecyl Sucrose, or 0.5% Tridecyl Maltoside onBlood Glucose Values in Rats Surfactant Agent Dodecyl Sucrose DecylMaltoside Tridecyl Maltoside Time (min) Blood Glucose Concentration(mg/dl) −20 266 249 255 −10 305 287 307 Insulin Eye Drops Added 0 351337 323 10 347 304 309 20 252 292 217 30 161 221 131 40 120 164 100 50105 138 87 60 114 114 107 70 113 104 115 80 104 110 79 90 86 120 85 100113 92 76 110 107 81 74 120 112 87 75 Glucagon Eye Drops Added 130 11195 82 140 143 99 121 150 202 132 148 160 247 157 173 170 242 171 162 180234 180 162 190 211 189 156

Example 6 Intranasal Administration of 0.25% TDM Plus Insulin DecreasesBlood Glucose Levels In Vivo

Intranasal administration of drugs or agents are possible in animalmodels e.g., mice and rats, although the nasal opening in is very small.In the experiments and results described herein, an anesthesia-inducedhyperglycemia model was used (described in Examples above).Hyperglycemic animals were induced by an intraperitoneal (IP) injectioncontaining xylazine-ketamine and blood glucose levels were monitoredover a period of time. Immediately after the xylazine-ketamineinjection, there was an increase in the blood glucose levels as shown inFIG. 2 (closed dark circles), and blood glucose levels were about 450mg/dl. The increase in blood glucose levels was attributed to theinhibition of pancreatic insulin secretion. Blood glucose levels peak toabout 482 mg/dl by 30 minutes after the xylazine-ketamine injection(FIG. 2). Then, at approximately 33 minutes after the xylazine-ketamineinjection, 6 μL of insulin (Humalog) in 0.25% tetradecylmaltoside (TDM;or Intravail A) was administered intranasally using a long thinmicropipette tip, and blood glucose levels were monitored at about 15minute intervals. After administration of the 0.25% TDM/insulincomposition, there was a rapid decrease in blood glucose levels,reaching a low of about 80 mg/dl at about the 60 minute time point, orabout 30 minutes after the insulin administration (FIG. 2). At about the75 minute time point, blood glucose levels gradually returned to thebaseline level in a normoglycemic mouse, or about 80-100 mg/dl.

The results above were compared with animals treated with insulin alone(same dosage), minus 0.25% TDM (FIG. 2, open circles). The insulin onlytreatment showed blood glucose levels do not start to decline until atabout the 120 minute time mark, or about 110 minutes after the insulinadministration. Further, the blood glucose levels observed in animalstreated with insulin alone never return to normoglycemic levels, as wasobserved in those animals receiving insulin plus 0.25% TDM (FIG. 2).

Thus, these results again demonstrate that compositions of the inventionconsisting of certain alkyl glycosides or alkyl saccharides plus a drug,e.g., insulin, effectively lower blood glucose levels, and that theseeffects are measurable shortly after administration of the drug.

Example 7 Intranasal Administration of 0.25% TDM (Intravail A)+Exendin-4Decreases Blood Glucose Levels In Vivo

The ob/ob mouse model was utilized for the studies described herein.Friedman, J. M., Nature 404, 632-634 (2000). All animals received anintraperitoneal (IP) injection of a bolus of 2 g/kg glucose for purposesof determining glucose tolerance. At time 0 the experimental animalswere given about 100 micrograms/kg of exendin-4/0.25% TDM (exendin-4from American Peptide) either as 10 μl of nasal drops (FIG. 3; closedtriangles), or by IP injection (FIG. 3; closed circles), or by and IPinjection of saline alone (no drug, no TDM; FIG. 3; open circles).Control animals were previously performed and received no drugs. Theresults of this study are shown in FIG. 3.

FIG. 3 shows that glucose tolerance of the animals were different sinceblood glucose levels vary at time 0 when the animals received theglucose bolus. Regardless, of the glucose tolerance level at time 0,immediately after injection of the glucose bolus, blood glucose levelsincreased in all three animals. The blood glucose level of the animalreceiving the IP injection of saline alone does not decrease as rapidlyas the experimental animals receiving the drug. Moreover, the animalreceiving the IP injection of saline alone never reached a normoglycemiclevel (FIG. 3, open circles). In contrast, the experimental animals,after administration of nasal drops of exendin-4/TDM, or IP injection ofexendin-4/TDM, showed a rapid and immediate decrease in blood glucoselevels.

Also exendin-4 administered about 15-30 minutes ahead of the glucosebolus (before time 0 in FIG. 3; data not shown) produced an even morepronounced lowering of blood glucose effect, because the absorption ofthe hormone takes a certain amount of time to be absorbed and to beactive. Thus, exendin-4 (or Exenatide) which is currently in humanclinical trials, when combined with alkyl glycosides of the invention,effectively treats a hyperglycemic condition by lowering the bloodglucose levels of the hyperglycemic subject.

Example 8 Alkyglycosides have Antibacterial Activity by ReducingBacterial Log Growth

The cultures of Candida albicans (ATCC No. 10231), Aspergillus niger(ATCC No. 16404), Escherichia coli (ATCC No. 8739), Pseudomonasaeruginosa (ATCC No. 9027), and Staphylococcus aureus (ATCC No. 6538)were obtained from American Type Culture Collection, 10801 UniversityBoulevard, Manassas, Va. 20110-2209. The viable microorganisms used inthe invention were not more than five passages removed from the originalATCC culture. As described herein, one passage is defined as thetransfer of organisms from an established culture to fresh medium andall transfers are counted.

Cultures received from the ATCC are resuscitated according to thedirections provided by the ATTC. Cells grown in broth were pelleted bycentrifugation, resuspended in 1/20th the volume of fresh maintenancebroth, and combined with an equal volume of 20% (v/v in water) sterileglycerol. Cells grown on agar were scraped from the surface into themaintenance broth also containing 10% glycerol broth. Small aliquots ofthe suspension were dispensed into sterile vials and the vials werestored in liquid nitrogen or in a mechanical freezer at a temperature nohigher than about −50° C. When a fresh seed-stock vial was required, itwas removed and used to inoculate a series of working stock cultures.These working stock cultures were then used periodically (each day inthe case of bacteria and yeast) to start the inoculum culture.

All media described herein should be tested for growth promotion usingthe microorganisms indicated above under Test Organisms.

To determine whether the alkyl saccharides of the invention inhibitgrowth or have antibacterial activity, the surface of a suitable volumeof solid agar medium was inoculated from a fresh revived stock cultureof each of the specified microorganisms. The culture conditions for theinoculum culture is substantially as described in Table IV. For example,suitable media can include but is not limited to, Soybean-Casein Digestor Sabouraud Dextrose Agar Medium. The bacterial and C. albicanscultures was harvested using sterile saline TS, by washing the surfacegrowth, collecting it in a suitable vessel, and adding sufficientsterile saline TS to obtain a microbial count of about 1×10⁸colony-forming units (cfu) per mL. To harvest the cells of A. niger, asterile saline TS containing 0.05% of polysorbate 80 was used, and thenadding sufficient sterile saline TS to obtain a count of about 1×10⁸ cfuper mL.

Alternatively, the stock culture organisms may be grown in any suitableliquid medium (e.g., Soybean-Casein Digest Broth or Sabouraud DextroseBroth) and the cells harvested by centrifugation, and washed andresuspended in sterile saline TS to obtain a microbial count of about1×10⁸ cfu per mL. The estimate of inoculum concentration was determinedby turbidimetric measurements for the challenge microorganisms. Thesuspension should be refrigerated if it is not used within 2 hours. Toconfirm the initial cfu per mL estimate, the number of cfu per mL ineach suspension was determined using the conditions of media andmicrobial recovery incubation times listed in Table IV (e.g., from about3 to about 7 days). This value serves to calibrate the size of inoculumused in the test. The bacterial and yeast suspensions were used within24 hours of harvest; whereas the fungal preparation can be stored underrefrigeration for up to 7 days.

TABLE IV Culture Conditions for Inoculum Preparation MicrobialIncubation Inoculum Recovery Temper- Incubation Incubation OrganismSuitable Medium ature Time Time Escherichia coli Soybean-Casein 32.5 ±18 to 24 3 to 5 (ATCC No. Digest Broth; 2.5 hours days 8739)Soybean-Casein Digest Agar Staphylococcus Soybean-Casein 32.5 ± 18 to 243 to 5 aureus Digest Broth; 2.5 hours days (ATCC No. Soybean-Casein6538) Digest Agar Candida albicans Sabouraud 22.5 ± 44 to 52 3 to 5(ATCC No. Dextrose Agar; 2.5 hours days 10231) Sabouraud Dextrose BrothAspergillus niger Sabouraud 22.5 ± 6 to 10 3 to 7 (ATCC No. DextroseAgar; 2.5 days days 16404) Sabouraud Dextrose Broth

To determine which alkylglycoside formulations have antibacterialactivity, the formulations were prepared in phosphate buffered saline(PBS) at pH 7. As a source of nutrition, either 1.5 mg/mL bovine serumalbumin (BSA; see Tables V and VI) or 1 mg/mL of PYY was added (seeTable VII) to the medium. BSA (CAS Number: 9048-46-8) was obtained fromSigma-Aldrich, St. Louis, Mo., USA,n-dodecyl-4-O-α-D-glucopyranosyl-β-D-glucopyranoside andn-tetradecyl-4-O-α-D-glucopyranosyl-β-D-glucopyranoside were obtainedfrom Anatrace Inc., Maumee, Ohio, USA, and PYY was obtained from BachemCalifornia Inc., Torrance, Calif., USA.

Antibacterial activity of the alkylglycosides were conducted in foursterile, capped bacteriological containers of suitable size into which asufficient volume of alkylglycoside solution had been transferred. Eachcontainer was inoculated with one of the prepared and standardizedinoculums and mixed. The volume of the suspension inoculum was betweenabout 0.5% and about 1.0% of the volume of the alkylglycoside solution.The concentrations of test microorganisms added to the alkylglycosidesolution was such that the final concentrations of the test preparationafter inoculation was between about 1×10⁵ and 1×10⁶ cfu per mL ofalkylglycoside solution. To determine the level of inhibition of growth,or reduction of growth based on a logarithmic scale, the initialconcentration of viable microorganisms in each test preparation wasestimated based on the concentration of microorganisms in each of thestandardized inoculum as determined by the plate-count method. Theinoculated containers were then incubated at about 22.5° C.±2.5. Thegrowth or non-growth of the microorganisms in each culture/containerwere again determined at day 14 and day 28. The number of cfu present ineach calculation was determined by the plate-count procedure standard inthe art for the applicable intervals. The change in the orders ofmagnitude of bacterium and/or fungi was then determined by subtractingthe first calculated log 10 values of the concentrations of cfu per mLpresent at the start or beginning (e.g., day 0), from the log 10 valuesof the concentration of cfu per mL for each microorganism at theapplicable test intervals (e.g., day 14 and day 28; see Tables V, VI andVII).

TABLE V Log reduction of microorganisms in cultures containing 0.125%n-Dodecyl-4-O-α-D-glucopyranosyl-β-D-glucopyranoside StaphlococcusEscherichia Candida Aspergillus aureus Coli albicans niger Day 0 7.3 ×10⁵ 1.2 × 10⁵ 3.2 × 10⁵ 4.8 × 10⁵ (cfu/gm) (cfu/gm) (cfu/gm) (cfu/gm)Day 14 ≥5.2 orders of N.D. 3.0 orders of 0.7 orders of magnitudemagnitude magnitude reduction reduction reduction Day 28 ≥5.2 orders of0.1 orders of ≥5.3 orders of No growth magnitude magnitude magnitudefrom initial reduction reduction reduction count

TABLE VI Log Reductions in cultures containing 0.2% n-Tetradecyl-4-O-α-D-glucopyranosyl-β-D-glucopyranoside Staphlococcus EscherichiaCandida Aspergillus aureus Coli albicans niger Day 0 7.3 × 10⁵ 1.2 × 10⁵3.2 × 10⁵ 4.8 × 10⁵ (cfu/gm) (cfu/gm) (cfu/gm) (cfu/gm) Day 14 ≥5 ordersof N.D. 3.0 orders of 0.5 orders of magnitude magnitude magnitudereduction reduction reduction Day 28 ≥5 orders of No growth ≥5.4 ordersof No growth magnitude from initial magnitude from initial reductioncount reduction count

Determining the antibacterial activity of other alkylglycosides wouldoccur substantially as described herein.

Example 9 Administration of Alkyglycosides with AntisenseOligonucleotides to Primates

TABLE VII Log reduction of cultures containing 0.25% n-Dodecyl-4-O-α-D-glucopyranosyl-β-D-glucopyranoside Staphlococcus EscherichiaCandida Aspergillus aureus Coli albicans niger Day 0 7.3 × 10⁵ 1.2 × 10⁵3.2 × 10⁵ 4.8 × 10⁵ (cfu/gm) (cfu/gm) (cfu/gm) (cfu/gm) Day 14 ≥4.9orders of ≥5 orders of ≥4.5 orders of 4.7 orders of magnitude magnitudemagnitude magnitude reduction reduction. reduction. reduction Day 28≥4.9 orders of ≥5 orders of ≥4.5 orders of 4.7 orders of magnitudemagnitude magnitude magnitude reduction reduction. reduction. reduction

An approximately 7,000 Dalton antisense oligonucleotide (ASO) with amodified backbone (phosphorothioate oligonucleotide as described in U.S.Pat. No. 7,132,530) mixed with alkylglycosidetetradecyl-beta-D-maltoside (Intravail™), was administered to sixCynomolgus monkeys canulated into the jejunum at a dose of 10 mg/kg. Theanimals were fasted prior to administration. Test agents were dissolvedin PBS buffer and injected through the cannula into the jejunum of eachanimal in a 1.5 mL volume or administered subcutaneously (s.c.) as notedin Table VIII.

TABLE VIII Bioavailability of Antisense Drugs Administered WithTetradecyl-Beta-D-Maltoside Average Bioavailability Test Agents (n = 6)Observations ASO (no tetradecyl- 0% Intestinal pili completelybeta-D-maltoside) (undetectable) intact intrajejunal ASO administereds.c. 100%  Intestinal pili completely at 0.5 mg/kg. intact 10 mg ASO +50 18% +/− 7%  Intestinal pili completely mg/kg tetradecyl- intactbeta-D-maltoside A5 intrajejunal 10 mg ASO + 9% +/− 7% The tops of someof the 50 mg/kg sodium intestinal pili were found caprate intrajejunalto be missing

The protocol involved a 3 way crossover in which each animal had thefirst 3 test agents in Table VIII administered on 3 different dates.There was a 1 week washout period between dosing dates. Two of theanimals were subsequently given a fourth test agent containing 5% sodiumcaprate as an absorption enhancer. Analysis of the blood levels wasconducted using quantitative analysis involving solid phase extractionusing cationic polystyrene nanoparticles.

Solid-phase extractions of the blood samples were first performed.Nanoparticle-oligonucleotide conjugates were formed using a known amountof oligonucleotide added to an aliquot of each sample (200-400 μl) anddiluted with 800 μl of 50 mM Tris.HCI (pH 9) in deionized water. Themixture was briefly vortexted prior to the addition of 200 μl of apolystyrene nanoparticle suspension prepared by surfactant-free emulsionpolymerization using water-soluble cationic initiators to induce apositive surface charge (solid content: approximately 10 mg/ml). Themixture was subsequently vortexed again. After 5-10 min of incubation,the suspension was centrifuged and the supernatant removed. Theparticles were resuspended in 1 ml of a solution of 0.5 M acetic acid indeionized water/ethanol (1:1) and separated from the washing solution bycentrifugation. After the supernatant was removed, the particles wereresuspended in 1 ml of deionized water and separated by anothercentrifugation step. 200 μl of a solution of 150 μM SDS in aqueousammonia (25%)/acetonitrile (60/40) was added to thenanoparticle-oligonucleotide conjugates and the releasedoligonucleotides were separated from the carrier by centrifugation. Inorder to exclude contamination of the samples with residual particles,the supernatant was placed in another 1.5-ml tube and centrifuged again.Subsequently, the samples were dried by rotoevaporation orlyophilization and stored at −20° C. until analysis.

Quantitative analysis was performed with capillary gel electrophoresisof the extracted samples. Capillary gel electrophoresis (CGE) wasperformed with a capillary electrophoresis system. An oligonucleotideanalysis kit containing polyvinyl alcohol (PVA) coated capillaries,polymer solution B, and oligonucleotide buffer was obtained. UsingPVA-coated capillaries, analysis was carried out using the manufacturesprotocol.

Using the data obtained from CGE analysis, quantitation ofphosphorothioate oligonucleotides was carried out. The amount ofoligonucleotides in the samples (n_(ON)) was calculated using thefollowing formula:n _(ON) =n _(Std)(ε_(Std)/ε_(ON))((A _(ON) /T _(ON))/(A _(Std) /T_(Std))),where n_(Std) is the amount of standard oligonucleotide added to thesample, ε_(std) and ε_(ON) are the molar extinction coefficients, andA_(Std) T_(Std) and A_(ON) T_(ON) are the corrected peak areas (quotientof peak area and migration time) of the standard and the investigatedcompound, respectively. The quotient of the corrected peak areas of theanalyte and the standard is referred to as the normalized area.

AUC's were calculated from the concentration vs. time cures over a 240minute period. The relative bioavailabilities were determined as theratio of each AUC divided by the AUC for the intravenously administereddrug. Intravail™ (tetradecyl-beta-D-maltoside) excipient providedbioavailability up to 18%. The control showed no detectable absorptionwithout a surfactant excipient. The sodium caprate formulation showed anaverage bioavailability of 9%.

Example 10 Preparation of Fast-Dispersing Dosage Forms of Olanzapine

Fast-dispersing dosage forms of olanzapine were prepared as follows.Olanzapine, CAS #132539-06-1, is obtained from SynFine (Ontario,Canada). Sodium acetate buffer, 10 mM, pH 5.0 and pH 6.5 is prepared asfollows. In an appropriate sized clean container with volumetricmarkings, place 495 mL of sterile water for injection. Add 0.286 mLacetic acid. Add 1N NaOH to bring the pH to 5.00 (or to pH 6.5). Whenthe proper pH is obtained, add additional water to bring the totalvolume to 500 mL and recheck the pH.

Liquid formulations having the compositions illustrated in Table IXbelow are made up by adding the fish gelatin or porcine skin gelatinslowly to the acetate buffer and allowing sufficient time to dissolvewhile stirring throughout the process. Upon complete dissolution of thefish or porcine skin gelatin, the mannitol is added and allowed todissolve. Then the sweetener is added. Once this has been fullydispersed, the active ingredient, olanzapine, being one of the examplesfor the compounds of the present invention, is added to produce thefinal solution. Secondary components such as preservatives,antioxidants, surfactants, viscosity enhancers, coloring agents,flavoring agents, sweeteners or taste-masking agents may also beincorporated into the composition. Suitable coloring agents may includered, black and yellow iron oxides and FD & C dyes such as FD & C blueNo. 2 and FD & C red No. 40 available from Ellis & Everard. Suitableflavoring agents may include mint, raspberry, licorice, orange, lemon,grapefruit, caramel, vanilla, cherry and grape flavors and combinationsof these. Suitable sweeteners include aspartame, acesulfame K andthaumatin. Suitable taste-masking agents include sodium bicarbonate.Cyclodextrins should be avoided since they form inclusion compounds withalkylsaccharides that reduce the effectiveness of these excipients.

Aliquots of 1 mL each of the above drug solutions are placed in thewells of a 24 well disposable microwell plastic plate. The micro wellplate containing the liquid aliquots is frozen at −70° in the frozenplate is placed within a glass lyophilization flask attached to aLabConco Freezone Model 4.5 desktop freeze drier and lyophilized undervacuum. Following lyophilization, the rapidly dispersing tablets arestored in the micro well plate in a dry environment until tested.Sucrose mixed mono- and di-stearate was provided as a gift by Croda Inc.and is designated CRODESTA F-110. Dodecyl maltoside, tetradecylmaltoside and sucrose mono-dodecanoate is obtained from Anatrace Inc.,Maumee, Ohio.

TABLE IX Olanzopine Formulations Example No. Ingredients 1 2 3 4 5 6 7Olanzapine¹  62.5 mg   125 mg   250 mg   500 mg   125 mg   250 mg   500mg Fish Gelatin ²  1.3 g  1.3 g  1.3 g  1.3 g — — — (3101) Dodecyl   250mg   500 mg — —   250 mg   500 mg — maltoside³ Sucrose — —   250 mg  750 mg — —   750 mg dodecanoate⁴ Gelatin Type A⁵ — — — —  1.3 g 1.52.0 Mannitol    1 g    1 g    1 g    1 g    1 g    1 g    1 g EP/USPAcesulfame K 0 .062 g 0 .062 g 0 .062 g 0 .062 g — — — Aspartame — — — —0.125 g 0.125 g 0.125 g Acetate Buffer Q_(s) Q_(s) Q_(s) Q_(s) Q_(s)Q_(s) Q_(s) (mL) 25 mL 25 mL 25 mL 25 mL 25 mL 25 mL 25 mL ¹SynFine,Ontario, Canada ² Croda Colloids Ltd (non-hydrolysed, spray dried fishgelatin) ⁴Sucrose dodecanoate (monoester)-Anatrace Inc. ⁵Sigma Aldrich(Gelatin Type A, porcine skin-G6144) Q_(s) = sufficient to give.

The drug olanzapine, also called Zyprexa, is known to be well absorbedwhen administered as a “whole-swallowed” tablet and reaches peakconcentrations in approximately 6 hours following an oral dose. It iseliminated extensively by first pass metabolism, with approximately 40%of the dose metabolized before reaching the systemic circulation.Pharmacokinetic studies showed that “whole-swallowed” olanzapine tabletsand rapidly dispersing olanzapine tablets prepared by lyophilization inthe manner described above in this Example, which disintegrate in about3 seconds to 10 seconds when placed in the mouth, are bioequivalent,exhibiting peak concentrations at about 6 hours after administration.Similarly, the first-pass effect in the liver eliminates approximately40% of the dose before reaching systemic circulation.

In the present example, fast-dispersing tablets are prepared bylyophilization as described above in this Example containing 10 mgolanzapine. Upon administration of the fast dispersing olanzapine tabletby placing it in contact with buccal tissue, it has been discovered thataddition of certain alkylsaccharides having specific alkyl chain lengthsto the fast-dispersing olanzapine tablets results in substantiallyreduced first-pass effect metabolism of olanzapine as seen by areduction in the relative proportion of olanzapine metabolites insystemic circulation compared to un-metabolized active drug. Therelative proportions of olanzapine and olanzapine metabolites in serumor plasma can be determined using an HPLC Chromatograph, Perkin Elmer200, with a Refractive Index Detector equipped with a thermostated cell.A suitable solid-phase absorbent may be used such as Lichrosorb RP-18(Merck, Darmstadt, Germany) 250 mm, with a mobile phase consisting ofacetonitrile:water gradient. Injection volumes of 20 μL using the PerkinElmer 200 auto-sampler and a flow rate of 0.8 mL/minute are satisfactoryfor this purpose. Specifically, incorporation of from 0.2% up to 10%dodecyl maltoside or tetradecyl maltoside or sucrose dodecanoate in afast-dispersing tablet format increases the drug that enters intosystemic circulation and decreases the drug that is eliminated by the“first-pass” effect in the liver. Additionally, the time to maximum druglevels is dramatically reduced, typically from one to six hours, to aslittle as approximately 15 to 45 minutes. For use in treating combativepatients undergoing psychotic episodes, this more rapid absorption ofdrug, resulting in more rapid onset of action, may be of great benefit.

Example 11 Preparation of Fast-Dispersing Dosage Forms of Melatonin

Melatonin or 5-methoxy-N-acetyltryptamine is a neurohormone used toregulate sleep-wake cycles in patients with sleep disorders. Endogenousmelatonin is secreted by the pineal gland in all animals exhibitingcircadian or circannual rhythms. Melatonin plays a proven role inmaintaining sleep-wake rhythms, and supplementation may help to regulatesleep disturbances that occur with jet lag, rotating shift-work,depression, and various neurological disabilities.

Commercially available formulations of melatonin include oral andsublingual tablets, capsules, teas, lozenges, and oral spray deliverysystems. Oral melatonin administration follows a differentpharmacokinetic profile than that of the endogenous hormone. After oraladministration, melatonin undergoes significant first-pass hepaticmetabolism to 6-sulfaoxymelatonin, producing a melatonin bioavailabilityestimated at 30-50%. DeMuro et al. (2000) reported that the absolutebioavailability of oral melatonin tablets studied in normal healthyvolunteers is somewhat lower at approximately 15%. The mean eliminationhalf-life of melatonin is roughly 45 minutes.

Fast-dispersing melatonin tablets are prepared containing 1 mg, 5 mg, 10mg and 20 mg according to the method described in Example 10 above, withand without 1% to 2% alkylsaccharide as described in Example 10. NewZealand White rabbits are anesthetized are placed into a restraining boxand anesthetized using a single administration of acepromazine/ketamine(0.7 mg/0.03 mg in 0.1 mL) administered by injection into the marginalear vein) to facilitate dosing. This results in anesthesia for a periodof about 10 minutes during which time the animals are dosed with testarticle. Thereafter, the animals return to consciousness. At individualtime point over a two hour period, 1 mL blood samples are collected fromthe central ear artery. After collection, plasma is immediately preparedfrom each blood sample using lithium/heparin as the anticoagulant. Allsamples are stored at −70° C. until assaying for melatonin. Melatonin ismeasured using a commercial ELISA kit manufactured by GenWay BiotechInc., San Diego, Calif. Upon administration by contacting thefast-disintegrating tablets with buccal tissue in the upper portion ofthe mouth, melatonin is found to be absorbed with a bioavailability ofat least 75% as measured by area under the curve in the presence ofalkylsaccharide and less than 50% in the absence of alkylsaccharide.Melatonin is measured using a commercial ELISA kit (No. 40-371-25005)manufactured by GenWay Biotech Inc., San Diego, Calif. In addition, forthe tablets containing alkylsaccharide the maximal concentration ofmelatonin is reached in approximately one half the time it takes fortablets not containing alkyl saccharides.

Example 12 Preparation of Fast-Dispersing Dosage Forms of Raloxifene

Raloxifene, also called Evista® is used for the treatment and preventionof osteoporosis in postmenopausal women, the reduction in the risk ofinvasive breast cancer in postmenopausal women with osteoporosis, andthe reduction in the risk of invasive breast cancer in postmenopausalwomen at high risk of invasive breast cancer. The recommended dosage isone 60 mg tablet daily. While approximately 60% of an oral dose ofraloxifene is absorbed rapidly after oral administration, presystemicglucuronide conjugation is extensive, resulting in an absolutebioavailability for raloxifene of only 2%. A fast-dispersing 60 mgraloxifene tablets prepared as described in U.S. Pat. Nos. 5,576,014 or6,696,085 B2 or 6,024,981 are found to have very similarpharmacokinetics with approximately 2% absolute bioavailability.However, a fast-dispersing tablet containing 10 mg or less of micronizedraloxifene—prepared by spray-dried dispersion (Bend Research Inc., BendOreg., or AzoPharma, Miramar, Fla.) or by more commonly used standardpharmaceutical grinding or milling processes, and 0.5% to 5% dodecylmaltoside, when administered buccally achieves systemic drug levelssimilar to those achieved with the 60 mg oral tablet and at the sametime results in less circulating inactive raloxifene glucuronide.

While clinical benefit results primarily from the unconjugated drug,side effects may be mediated by either or both active drug andsubstantially inactive glucuronide conjugated drug. Thus reducingexposure to the inactive drug conjugate, in this case present in as muchas a 30-fold higher concentration than active drug, affords potentiallysignificant clinical benefit in reducing the likelihood of side effects.Raloxifene has a water solubility of approx. 0.25 mg/L. As a result, itis not possible to dissolve raloxifene in water in preparation forlyophilization to prepare a fast-dispersing formulation as described inExample 10.

In this case, a self-assembling hydrogel can be formed by adding 1% to30% w/w CRODESTA F-110 in a suitable buffer, which is vortexed andheated to 45 degrees for 1 hr. Then raloxifene in a fine particle ormicronized form is added to the warm liquid to achieve a concentrationin suspension of 60 mg/mL which is again mixed by vortexing until thesolid is uniformly suspended and dispersed. Upon cooling to roomtemperature, a stable thixotropic hydrogel forms which is capable ofbeing dispensed but which maintains the uniform suspension. Acetatebuffer in the pH range of pH 2 to pH 7 is found to be particularly wellsuited for this purpose. Aliquots of 1 mL of the gel suspension ofraloxifene are placed in the wells of a 24 well disposable microwellplastic plate and lyophilized as described in Example 1.

Administration of this fast dispersing formulation upon presentation tobuccal tissue results in an increase (a doubling) in absolutebioavailabilty to at least 4% and a corresponding measurable reductionin the ratio of circulating raloxifene glucuronide conjugateconcentration to unconjugated raloxifene.

Example 13 Preparation of Fast-Dispersing Dosage Forms ofDiphenhydramine

Diphenhydramine is a sedating antihistamine with pronounced centralsedative properties and is used as a hypnotic in the short-termmanagement of insomnia, symptomatic relief of allergic conditionsincluding urticaria and angioedema, rhinitis and conjunctivitis,pruritic skin disorders, nausea and vomiting, prevention and treatmentof motion sickness, vertigo, involuntary movements due to the sideeffects of certain psychiatric drugs and in the control of parkinsonismdue to its antimuscarinic properties. A particularly desirablecharacteristic of diphenhydramine is its apparent lack of any evidenceof creating dependency. Because of its excellent safety profile, it isavailable as an over-the-counter drug and unlike some of the newer sleepmedications such as Ambien® and Lunesta® which can cause bizarrebehaviors such as sleepwalking and eating-binges while asleep, alongwith occasional severe allergic reactions and facial swelling causingthe FDA to require label warnings about these side effects for thesenewer prescription medications.

Diphenhydramine hydrochloride is given by mouth in usual doses of 25 to50 mg three or four times daily. The maximum dose in adults and childrenis about 300 mg daily. A dose of 20 to 50 mg may be used as a hypnoticin adults and children over 12 years old. The drug is well absorbed fromthe gastrointestinal tract; however it is subject to high first-passmetabolism which appears to affect systemic drug levels. Peak plasmaconcentrations are achieved about 1 to 4 hours after oral doses.Diphenhydramine is widely distributed throughout the body including theCNS and due to its extensive metabolism in the liver, the drug isexcreted mainly in the urine as metabolites with small amounts ofunchanged drug found to be present.

While diphenhydramine is considered safe and effective for treatment ofinsomnia and other disorders, the relatively long onset of action due tothe delay in achievement of peak plasma concentrations of from one tofour hours is inconvenient and reduces the practical utility of thissafe and effective drug. Intravenously administered diphenhydramineexerts a rapid onset of action; however, intravenous administration isnot practical for outpatient use or non-serious medical indications. Theneed for a rapid onset-of-action formulation of diphenhydramine isclear. In the case of insomnia, a patient may need to take the currentoral forms of the drug well in advance of going to bed in order tominimize the likelihood of extended restless sleeplessness while waitingfor the drug to achieve sufficient systemic drug levels in order toexert its desired pharmacological effect. In the case of the antiemeticapplications of diphenhydramine, rapid onset of action is also highlydesirable in order to relieve nausea and vomiting as soon as quickly aspossible. This is likewise the case in the treatment of motion sicknessand vertigo since these symptoms can arise unexpectedly and it is bothinconvenient and undesirable to have to wait one to four hours while theorally administered drug achieves sufficient systemic drug levels toachieve its beneficial effects.

Diphenhydramine has a solubility in water of approximately 3.06 mg/mL.Therefore the method described in Example 12 may be used to preparefast-dispersing diphenhydramine tablet containing 50 mg of drug and 1%to 2% alkylsaccharide. Because Diphenhydramine is slightly bitter, ataste masking amount of a pharmaceutically acceptable flavor and asweetener may be added to improve palatability. Fast-dispersing tabletsprepared in this manner have a more rapid onset of action compared to“whole-swallowed” tablets syrup, chewable tablets, lozenge, or ediblefilm-strip and exhibit less first-pass metabolism as well.

Example 14 Administration of Alkylglycosides with Anti-Obesity PeptideMouse [D-Leu-4]OB3 to Mice

This example shows the uptake of anti-obesity peptide mouse [D-Leu-4]OB3in 0.3% alkylglycoside tetradecyl-beta-D-maltoside (Intravail™ A3) bymale Swiss Webster Mice. The synthetic leptin agonist [D-Leu-4]OB3 mixedwith 0.3% alkylglycoside tetradecyl-beta-D-maltoside (Intravail™ A3),was administered to six-week old male Swiss Webster mice at a dose of 1mg by gavage.

Mouse [D-Leu-4]OB3 (at a concentration of 1 mg/200 ul) was dissolved ineither PBS (pH 7.2) or 0.3% alkylglycoside tetradecyl-beta-D-maltoside(Intravail™ A3) reconstituted in PBS (pH 7.2) and administered bygavage, without anesthesia, to each of 4 mice per time point. After 10,30, 50, 70, 90 or 120 minutes, the mice were euthanized by inhalation ofisoflurane (5%) and exsanguinated by puncture of the caudal vena cava.Blood was also collected from four mice not given peptide (prebleed).The blood from each of the four mice in the time period was pooled, andserum samples were prepared. Mouse [D-Leu-4]OB3 content of the pooledsamples was measured by competitive ELISA.

These experiments were repeated twice. The data collected from a singleexperiment are presented in Table X and FIG. 4. The data were determinedto be highly reproducible. Uptake curves were plotted using Microsoft™Excel, and AUC was calculated using a function of the graphics programSigmaPlot 8.0™ (SPSS Science, Chicage, Ill.). The lowest AUC valueobtained was arbitrarily set at 1.0. Relative bioavailability wasdetermined by comparing all other AUC values to 1.0.

TABLE X Uptake of 1 mg Mouse p-Leu-4]OB3 in 0.3% AlkylglycosideTetradecyl-beta-D-maltoside (Intravail ™ A3) By Male Swiss Webster MiceFollowing Administration By Gavage Sample AUC Relative bioavailabilityMouse [D-Leu-4]OB3 137,585 ng/ml/min 1.0 in PBS Mouse [D-Leu-4]OB3552,710 ng/ml/min 4.0 in 0.3% alkylglycoside tetradecyl-beta-D-maltoside (Intravail ™ A3)

As evidenced in Table X and FIG. 4, addition of alkylglycosidetetradecyl-beta-D-maltoside (Intravail™ A3) at 0.3% increases relativeabsorption of the OB-3 peptide by 4-fold compared to peptide in PBSalone.

Example 15 Administration of Alkylglycosides with Sumatriptan to Canines

This example shows the uptake of sumatriptan in 0.5% alkylglycosidetetradecyl-beta-D-maltoside (Intravail™ A3) by canines. Sumatriptanmixed with 0.5% alkylglycoside tetradecyl-beta-D-maltoside (Intravail™A3), was administered to canines as a dose of 25 mg by both oral andrectal administration.

As evidenced in FIG. 5, addition of alkylglycosidetetradecyl-beta-D-maltoside (Intravail™ A3) at 0.5% increases C_(max) ofsumatriptan for both oral and rectal administration as compared tocurrently available 25 mg oral tablets. C_(max) for currently availabletablets was determined to be 104 ng/ml for canines as represented by thehorizontal dashed line in FIG. 5.

Example 16 Oral Administration of Octreotide

Octreotide in three oral concentrations of n-dodecyl-beta-D-maltoside(DDM) (0.5%, 1.5%, and 3% DDM) and a subcutaneous injection (s.c.) ofoctreotide in buffer (s.c. Octreotide) containing no Intravail® isadministered to four respective groups of 24 mice each. The animal testgroups are described below in Table 2. Dosing solutions may be storedrefrigerated at 4-8 deg. C. prior to administration. The oral andsubcutaneous doses administered are adjusted to be 1000 μg/kg averagebody mass/group (which is 30 μg for 30 g mice), administered by 200 μLoral gavage or subcutaneously as a 100 uL injection between the skin andunderlying tissue layers in the scapular region on the back of eachanimal. Mice are anesthetized with 5% isoflurane, and blood is collectedby cardiac puncture over a three hour time period at 0, 5, 10, 15, 30,60, 120 and 180 minutes immediately prior to or following either oral orsubcutaneous administration of octreotide. Death is confirmed bycervical dislocation. After blood collection, serum is immediatelyprepared from each blood sample. Dosing solutions and all serum samplesare stored at −70° C. until assayed as described below. Collection ofblood and serum preparation—5, 10, 15, 30, 60, 120 or 180 min afteroctreotide delivery, the mice (n=six per time point) are anesthetizedwith isoflurane (5%) and exsanguinated by cardiac puncture. Euthanasiais confirmed by cervical dislocation. The blood is collected in sterilenonheparinized plastic centrifuge tubes and allowed to stand at roomtemperature for 1 h. The clotted blood is rimmed from the walls of thetubes with sterile wooden applicator sticks. Individual serum samplesare prepared by centrifugation for 30 min at 2600×g in an Eppendorf5702R, A-4-38 rotor (Eppendorf North America, Westbury, N.Y., USA), Theserum samples in each experimental group are pooled and stored frozenuntil assayed for octreotide content by EIA. The three treatment groupsand s.c. control group are as follows: 1) oral Octreotide, 0.5% DDM; 2)oral Octreotide 1.5% DDM; 3) oral Octreotide, 3.0% DDM; and 4) s.c.Octreotide. At time zero (0), octreotide is delivered subcutaneously orby gavage to each mouse. Following treatment, the mice are transferredto separate cages for the designated time period.

The Animal Test System that was used in these studies is described inTable XI below. Animals are segregated by weight into each of the fourtreatment groups to minimize variation within groups. The animals arehoused individually in polycarbonate cages fitted with stainless steelwire lids and air filters and supported on ventilated racks (ThorenCaging Systems, Hazelton, Pa., USA). The mice are maintained at aconstant temperature (24° C.) with lights on from 07:00 to 19:00 hoursand allowed food and water ad libitum.

TABLE XI Animal Test System Species (strain): Male Swiss Webster (SW-M)mice, 6 to 7 weeks of age Supplier: Taconic Farms # of males: Fourgroups of 24 mice (96 total) # of females 0 Age: 6 to 7 weeks of ageHousing: Three animals in plastic shoebox cages Food: Rodent ChowAvailability of water: Ad lib Availability of food: Ad lib

Octreotide is obtained from BCN (Spain) or Polypeptide Laboratories(California, USA). Octreotide stock solutions are prepared as describedin Table XII by dissolving the lyophilized powder in pH 4.5 acetatebuffer 0.1% EDTA (Table 2) containing 0.0% DDM (s.c. control), 0.5%,1.5% or 3% DDM. The appropriate dose is administered to animals in eachgroup as listed in Table XIII. All of these animal procedures arereviewed and approved by the institutional Animal Care and UseCommittee, and are performed in accordance with relevant guidelines andregulations. The dosing solution remaining after administration isdivided and frozen at −70° C. until assayed.

TABLE XII pH 4.5 mM Sodium Acetate Buffer, 0.1% EDTA Component QuantityAcetic acid 0.286 mL 1N NaOH adjust to pH 4.5 Na2 EDTA 500 mg water 500mL Adjust pH: pH 4.5

TABLE XIII Dosing Solutions in pH 4.5 Acetate Buffer, 0.1% EDTA & DoseAdministration Total DDM Final Volume Dose (mg/ Octreotide OctreotideAdmin- (30 g Group 5 mL) (mg)* Concentration istered mouse) Oral 50 mgplus 1.5 mg in 150 ug/mL 200 μL 30 μg Octreotide- in 5 mL 5 mL (10 mLtotal 0.5% A3 vol.) Oral 150 mg plus 1.5 mg in 150 ug 200 μL 30 μgOctreotide- in 5 mL 5 mL (10 mL total 1.5% A3 vol.)/mL Oral 300 mg plus1.5 mg in 150 ug/mL 200 μL 30 μg Octreotide- in 5 mL 5 mL (10 mL total3% A3 vol.) s.c. N/A N/A 1.5 mg in 300 ug/mL 100 μL 30 μg Octreotide 5mL (5 mL total vol.) *Prepared as 6 mg dissolved in 20 mL acetate buffer

Octreotide concentrations for dosing solution(s) and pooled serumsamples for each time period for each treatment group are assayed intriplicate using an octreotide enzyme immunoassay assay (EIA) (PeninsulaLaboratories, LLC (San Carlos, Calif.) Cat. No. S-1342-Octreotide forSerum and Plasma Samples) according to the instructions supplied by themanufacturer.

Pharmacokinetic analyses is carried out as follows. To determinerelative bioavailability, serum concentrations of octreotide vs. timefollowing s.c. and oral delivery are plotted using the graphics programSigmaPlot™ 8.0 (SPSS Science, Chicago, Ill., USA). The area under eachcurve (AUC) is calculated with a function of this program. The lowestAUC value obtained is arbitrarily set at 1.0. Relative bioavailabilityis determined by comparing all other AUC values to 1.0.

Serum half-life (t _(1/2)) is determined as follows. The period of timerequired for the serum concentration of octreotide to be reduced toexactly one-half of the maximum concentration achieved following s.c. ororal administration is calculated using the following formula:t _(1/2)=0.693/k _(elim)

k_(elim) represents the elimination constant, determined by plotting thenatural log of each of the concentration points in the beta phase of theuptake profiles against time. Linear regression analysis of these plotsresults in straight lines, the slope of which correlates to the k_(elim)for each delivery method.

Clearance of octreotide from the plasma following s.c. or oral deliveryis calculated from the AUC using the following equation:CL=Dose/AUC

Since the half-life of a drug is inversely related to its clearance fromthe plasma and directly proportional to its volume of distribution, theapparent volume of distribution of octreotide following s.c. or oraldelivery is calculated from its half-life and clearance using thefollowing equation:t _(1/2)=(0:693×V _(d))/CL

Results: Octreotide uptake profiles following s.c. and oral delivery in0.5%, 1.5% or 3.0% Intravail® are shown in FIGS. 6 to 9, respectively.All of these profiles show biphasic uptake of octreotide with an initialpeak (C_(max1)) at 10 min (t_(max1)) followed by a second peak(C_(max2)) at 30 min (t_(max2)). C_(max1) and C_(max 2) areapproximately the same (6.67 ng/ml vs. 7.59 ng/ml, respectively)following s.c. administration, and decrease at different rates aftereach of the two peaks (FIG. 6).

Oral delivery of octreotide in 0.5% Intravail® produces an uptakeprofile (FIG. 7) with a C_(max1) more than 2-fold higher than C_(max2)(59.7 ng/ml vs. 25.9 ng/ml, respectively). When the Intravail®concentration is increased to 1.5% or 3.0%, C_(max1) is reduced to 17.8ng/ml and 3.75 ng/ml, respectively. Likewise 1.5% or 3.0% Intravail®also reduces C_(max2) to 4.0 ng/ml and 2.48 ng/ml, respectively. Asobserved after s.c. delivery, octreotide concentrations following oraldelivery in 0.5%, 1.5% or 3.0% Intravail® decrease at different ratesafter each of the two peaks.

The relative bioavailability of octreotide is determined by measuringthe area under the uptake curve (AUC) for each delivery method. Thisvalue represents the total extent of peptide absorption into thesystemic circulation, or total uptake, following its administration.Because of the biphasic nature of the uptake profiles, the relativebioavailability of octreotide following s.c. and oral delivery isdetermined by measuring the AUC for each of the two peaks in the profileseparately, and determined as follows: AUC=AUC₁+AUC₂. Using thisformula, the AUC of octreotide after s.c. administration is determinedto be 290 ng/ml/min, and assigned a relative bioavailability of 1.0. TheAUC of octreotide following oral delivery in 0.5%, 1.5% or 3.0%Intravail® is 1,254 ng/ml/min, 230.7 ng/ml/min, and 141.24 ng/ml,respectively, and is assigned relative bioavailabilities of 4.3, 0.8 and0.6.

To determine the serum half-life of octreotide following s.c. and oraldelivery, the k_(elim) for each peak in the uptake curves is calculatedseparately (k_(elim1) and k_(elim2)). These values are then used todetermine the half-life of octreotide under each peak (t_(1/21) andt_(1/2 2)). The overall half-life is calculated as follows:t_(1/2)=t_(1/2 1)+t_(1/2 2), and is determined to be 41.2 min followings.c. delivery and 53.1 min, 25.8 min and 23.6 min following oraldelivery in 0.5%, 1.5% or 3.0% Intravail®, respectively.

Because of the biphasic profile of the uptake curves associated withs.c. and oral delivery of octreotide, plasma CL is measured using theAUC associated with each peak in the profile: CL₁=Dose/AUC1 andCL₂=Dose/AUC₂. Overall clearance is calculated as follows: CL=CL₁+CL₂,and is determined to be 30 L/min following s.c. administration, and 3.9L/min, 28.4 L/min and 54.9 L/min following oral delivery in 0.5%, 1.5%or 3.0% Intravail®, respectively.

The apparent volume of distribution (Vd) of octreotide following s.c.and oral delivery is calculated using the half-life and clearance ratedetermined for each peak associated with the biphasic uptake profiles:t_(1/2 1)=0.693V_(d1)/CL₁ and t_(1/2 2)=0.693×V_(d2)/CL₂. The overallapparent volume of distribution is calculated as follows:V_(d)=V_(d1)+V_(d2) and determined to be 301.1 L following s.c. deliveryand 84.7 L, 299.3 L and 357.8 L following oral delivery in 0.5%, 1.5% or3.0% Intravail®.

When the pharmacokinetics of orally delivered (by gavage) octreotide inincreasing concentrations (0.5%, 1.5% and 3.0%) of DDM are compared tothe pharmacokinetics of octreotide delivered subcutaneously, oraldelivery of octreotide in 0.5% DDM is seen to significantly enhancedtotal uptake (1,254.08 ng/ml/min vs. 290.12 ng/ml/min, respectively) andrelative bioavailability (4.3 vs. 1.0, respectively) when compared todelivery by s.c. injection. Higher concentrations of DDM do not furtherenhance uptake or bioavailability. The half-life of octreotide isincreased by oral delivery in 0.5% DDM from 41.2 min (s.c.) to 52.1 min,and clearance from the plasma is reduced from 30.0 L/min (s.c.) to 3.9L/min. The results indicate that oral delivery of octreotide incompositions containing DDM is feasible, and is an effective method ofadministration for achieving high serum levels of octreotide whencompared to s.c. injection. All pharmacokinetic parameters measured inthis study are summarized in Table XIV. In addition to DDM, n-tetradecylmaltoside, n-tridecyl maltoside and sucrose monododecanoate may be usedto substitute for DDM to get similar results.

TABLE XIV Pharmacokinetic Parameters of Octreotide Uptake in Male SwissWebster Mice Following Subcutaneous Delivery in 10 mM Sodium AcetateBuffer Containing 0.1% EDTA (pH 4.5) or Oral Administration (by gavage)in Increasing Concentrations of DDM. Oral Oral Oral 0.5% 1.5% 3.0% S.C.DDM DDM DDM Cmax (ng/ml) C_(max1) 6.67 59.68 17.8 3.75 C_(max2) 7.5925.92 4.0 2.48 Tmax (min) T_(max1) 10 10 10 10 T_(max2) 30 30 30 30 AUC(ng/ml/min) 290.12 1254.08 230.70 141.24 AUC₁ 38.39 353.03 97.80 21.50AUC₂ 251.73 901.04 132.90 119.74 Relative bioavailability 1.0 4.3 0.80.6 kelim (ml/min) k_(elim1) 0.3400 0.4400 0.7200 0.5200 k_(elim2)0.0177 0.0132 0.0279 0.0311 t_(1/2) (min) 41.2 52.1 25.8 23.6 t_(1/21)2.04 1.56 0.96 1.33 t_(1/22) 39.15 52.50 24.84 22.28 CL (L/min) 30.0 3.928.4 54.9 CL₁ 26.0 2.8 20.9 46.5 CL₂ 4.0 1.1 7.5 8.4 Vd (L) 301.1 84.7299.3 357.8 V_(d1) 76.7 0.6 29.0 89.3 V_(d2) 224.4 84.1 270.3 268.5

Example 17 Oral Administration of the GLP-1 Analog, Liraglutide

Objective: Test oral bioavailability of liraglutide with Intravail™ A3during a challenge with dietary sugar

Procedure was as follows. 500 uL of liraglutide was extracted from pensand placed in 50 mm test tube. 50 uL 5% A3 was added in H2O-0.45% finalA3 concentration. Composition was mixed with disposable plastic dropper.40 grams of dietary sugar was ingested and 60 minutes allowed to pass.Blood glucose was measured (upper arm just above elbow on left arm)using meter=T_(o). Deposit composition on back of tongue and swallow.Blood glucose level was measured and recorded at T=10, 20, 30, 40, 55,70, 80 and 90 min.

Results: Blood glucose declined from 146 mg/dL down to 100 mg/dL at 80min. The rate of decline in he presence of a dietary sugar challenge wasslower than in the non-challenged state observed previously.

Conclusion: Oral liraglutide using Intravail A3 at 0.45% effectivelyincreases bioavailability and delivers liraglutide to the blood stream.The 500 uL dose was 1.4× the standard injectable dose and the PD effectwas large, so bioavailability is substantial—approximately 30% to 60%.This confirms previously observed high relative bioavailability ascompared to delivery without alkylglycoside. Effectiveness underconditions of dietary sugar challenge is also demonstrated. Bloodglucose levels are shown in FIG. 10 while the dose response is shown inFIG. 11. A positive dose response is seen further confirming thepharmacodynamics.

TABLE XV Effect of Orally Delivered Liraglutide on Blood Glucose ValuesLiraglutide Oral Data glucose (mg/dL) time Trial 1 Trial 2 −20 125 −10144 0 116 146 10 122 127 20 102 124 30 101 128 40 104 115 55 111 70 10880 100 90 105

Example 18 Oral Delivery of Phenylephrine

In a canine model, subjects were dosed orally with various phenylephrine(PE) tablet formulations, and blood samples were taken to determine therate of parent phenyl ethyl phenylephrine absorption. Blood levels ofparent phenylephrine were measured. Three subject groups were createdwith each alkylsaccharide test group containing five subjects in eachtest group. All subject groups received 10 mg of phenylephrine as asingle tablet. Each subject was in the fasting state for overnight andtwo hours post dose. Blood samples were taken at pre-dose, 5, 10, 15,30, 45 min., 1, 2, and 3 hours.

The tablet formulations tested are shown in Table XVI. Comparator wasthe standard 30 mg (total weight including excipients) phenylephrinetablet was no absorption enhancer. Group number 2 tablets contain thealkylsaccharide designated A3, which is n-dodecyl-beta-D-maltoside inthe amounts of 2.5 mg, 5 mg, and 7.5 mg, respectively. Group numberthree tablets contain the alkylsaccharide designated B3, which issucrose monododecanoate, in the amounts of 5 mg, 10 mg, and 20 mg,respectively.

TABLE XVI Comparator (1) Regular 10 mg PE Aegis A3 (2) 10 mg PE + 2.5 mgA3 10 mg PE + 5 mg A3 10 mg PE + 7.5 mg A3 Aegis B3 (3) 10 mg PE + 5 mgB3 10 mg PE + 10 mg B3 10 mg PE + 20 mg B3

TABLE XVII AUC (0-3 hour) Test Articles AUC (0-3 hour) Change Ratio 2.5mg A3 + 10 mg PE tablet 2229610 1.14   5 mg A3 + 10 mg PE tablet 30551401.56 7.5 mg A3 + 10 mg PE tablet 1648846 0.84

TABLE XVIII AUC (0-3 hour) Test Articles AUC (0-3 hour) Change Ratio  5mg B3 + 10 mg PE tablet 1684238 0.86 10 mg B3 + 10 mg PE tablet 29323601.50 20 mg B3 + 10 mg PE tablet 2333966 1.19

Results: As can be seen from FIG. 12 and Table XVII,n-dodecyl-beta-D-maltoside increased oral bioavailability ofphenylephrine in compressed tablets by up to 56%. Similarly, as can beseen from FIG. 13 and Table XVIII sucrose monododecanoate increased oralbioavailability of phenylephrine in compressed tablets by up to 50%.Interestingly, and unexpectedly, the maximum increase in bioavailabilityoccurred at intermediate levels, between the low and highalkylsaccharide levels tested. In the case ofn-dodecyl-beta-D-maltoside, this occurred at a loading a 5 mg ofalkylsaccharide. Whereas in the case of sucrose mono-dodecanoate thisoccurred at a loading of 10 mg of alkylsaccharide. However, a loweralkylsaccharide loading of dodecyl-beta-D-maltoside also showed anincrease over the comparator, whereas in the case of sucrosemono-dodecanoate, a slightly higher alkylsaccharide loading gave rise toan increase in bioavailability over the comparator. The loading ofalkylsaccharides in both cases has bracketed the relative amounts usefulin providing maximum bioavailability enhancement. The actual amounts mayvary as the amount of drug varies and as the choice of drug may alsovary. Thus, similar formulation studies may be conducted with otherdrugs intended to be formulated in solid dosage forms having enhancedbioavailability, such that the relative loadings of alkylsaccharide forthe desired drug dose can be individually determined. Other drugs forwhich these formulations and methods of testing are useful include byway of example phenylephrine (as HCl or bitartrate), aspirin, naproxensodium, brompheniramine (maleate), triprolidine, chlorpheniramine(maleate), dextromethorphan (HBr), guaifenesin, acetaminophen,pseudaphedrine, epinephrine, diphenhydramine, cimetidine, loratadine,ranitidine, famotidine, ketoprofen, omeprazole, clemastine,dimenhydrinate, ibuprofen, cyclizine (Marizine) or otherpharmaceutically acceptable salts thereof such as hydrochlorides,maleates, tartrates, acetates, and the like, as individual (monotherapy)drugs or as combinations of two or more drug substances in a soliddosage form.

Example 19 Administration of Alkylglycosides with Triptans IncreasesBioavailability

Sumatriptan sulfate, naratriptan-HCl; or rizatriptan benzoate isdissolved in 20 mM sodium acetate buffer, pH 5.5 containing 0%, 0.02%,0.05%, 0.1%, 0.2%, or 1.0% alkylsaccharide. Each set of drug solutionsis administered to six groups of eight rats each by 20 uL instillationto a single nare of each animal. 200 μl blood samples are drawn byorbital bleed over a three hour time period at 0, 5, 10, 15, 30, 60, 120and 180 minutes. After the last blood sample is collected each animal iseuthanized with CO₂. After blood collection, plasma is immediatelyprepared from each blood sample using lithium/heparin as theanticoagulant. Plasma samples are stored at −70° C. until analyzed.Plasma drug levels are determined by HPLC using the method described byBoulton or similar HPLC method. The concentration vs time data areplotted to determine the Cmax at each alkylsaccharide concentration andthe ratio of Cmax with and without alkylsaccharide present is calculatedand recorded as shown in Table XIX. The observed Tmax values from eachplot is determined by inspection of the concentration vs time plots andrecorded as shown in Table XX. The doses designated in this tablereflect amounts of triptan free base in each case.

TABLE XIX Cmax Ratio of Triptan Analog Administration Sumatriptan (6.2mg/kg dose) Ratio Cmax(alkylsacch)/Cmax(No alkylsacch) Alkylsaccharide2% 0.5% 0.1% 0.05% 0.02% octyl maltoside ≤1.5 ≤1.5 ≤1.5 ≤1.5 ≤1.5 decylmaltoside ≥1.5 ≥1.5 ≥1.5 ≤1.5 ≤1.5 dodecyl maltoside ≥1.5 ≥1.5 ≥1.5 ≥1.5≤1.5 tridecyl maltoside ≥1.5 ≥1.5 ≥1.5 ≥1.5 ≤1.5 tetradecyl maltoside≥1.5 ≥1.5 ≥1.5 ≥1.5 ≤1.5 dodecyl sucrose ≥1.5 ≥1.5 ≥1.5 ≥1.5 ≤1.5Naratriptan (0.22 mg/kg dose) Ratio Cmax(alkylsacch)/Cmax(No alkylsacch)Alkylsaccharide 2% 0.5% 0.1% 0.05% 0.02% octyl maltoside ≤1.5 ≤1.5 ≤1.5≤1.5 ≤1.5 decyl maltoside ≥1.5 ≥1.5 ≥1.5 ≤1.5 ≤1.5 dodecyl maltoside≥1.5 ≥1.5 ≥1.5 ≥1.5 ≤1.5 tridecyl maltoside ≥1.5 ≥1.5 ≥1.5 ≥1.5 ≤1.5tetradecyl maltoside ≥1.5 ≥1.5 ≥1.5 ≥1.5 ≤1.5 dodecyl sucrose ≥1.5 ≥1.5≥1.5 ≥1.5 ≤1.5 Rizatriptan (0.88 mg/kg dose) RatioCmax(alkylsacch)/Cmax(No alkylsacch) Alkylsaccharide 2% 0.5% 0.1% 0.05%0.02% octyl maltoside ≤1.5 ≤1.5 ≤1.5 ≤1.5 ≤1.5 decyl maltoside ≥1.5 ≥1.5≥1.5 ≤1.5 ≤1.5 dodecyl maltoside ≥1.5 ≥1.5 ≥1.5 ≥1.5 ≤1.5 tridecylmaltoside ≥1.5 ≥1.5 ≥1.5 ≥1.5 ≤1.5 tetradecyl maltoside ≥1.5 ≥1.5 ≥1.5≥1.5 ≤1.5 dodecyl sucrose ≥1.5 ≥1.5 ≥1.5 ≥1.5 ≤1.5

TABLE XX Tmax of Triptan Analog Administration Sumatriptan (6.2 mg/kgdose) Tmax (minutes) Alkylsaccharide 2% 0.5% 0.1% 0.05% 0.02% octylmaltoside ~60-120 ~60-120 ~60-120 ~60-120 ~60-120 decyl maltoside ~5-15~5-15 ~60-120 ~60-120 ~60-120 dodecyl maltoside ~5-15 ~5-15 ~5-15 ~5-15~60-120 tridecyl maltoside ~5-15 ~5-15 ~5-15 ~5-15 ~60 tetradecylmaltoside ~5-15 ~5-15 ~5-15 ~5-15 ~60 dodecyl sucrose ~5-15 ~5-15 ~5-15~5-15 ~60 Naratriptan (0.22 mg/kg dose) Tmax (minutes) Alkylsaccharide2% 0.5% 0.1% 0.05% 0.02% octyl maltoside ~60-120 ~60-120 ~60-120 ~60-120~60-120 decyl maltoside ~5-15 ~5-15 ~60-120 ~60-120 ~60-120 dodecylmaltoside ~5-15 ~5-15 ~5-15 ~5-15 ~60-120 tridecyl maltoside ~5-15 ~5-15~5-15 ~5-15 ~60 tetradecyl maltoside ~5-15 ~5-15 ~5-15 ~5-15 ~60 dodecylsucrose ~5-15 ~5-15 ~5-15 ~5-15 ~60 Rizatriptan (0.88 mg/kg dose) Tmax(minutes) Alkylsaccharide 2% 0.5% 0.1% 0.05% 0.02% octyl maltoside~60-120 ~60-120 ~60-120 ~60-120 ~60-120 decyl maltoside ~5-15 ~5-15~60-120 ~60-120 ~60-120 dodecyl maltoside ~5-15 ~5-15 ~5-15 ~5-15~60-120 tridecyl maltoside ~5-15 ~5-15 ~5-15 ~5-15 ~60 tetradecylmaltoside ~5-15 ~5-15 ~5-15 ~5-15 ~60 dodecyl sucrose ~5-15 ~5-15 ~5-15~5-15 ~60

Example 20 Nasal Administration of Sumatriptan with AlkylglycosideIncreases Bioavailability and C_(MAX) and Speeds Onset of SystemicAbsorption

Sumatriptan sulfate is dissolved in phosphate buffer prepared bydissolving 0.2 g dibasic sodium phosphate and 10.0 g monobasic potassiumphosphate in 1 L of water., adjusted to pH 5.5, containing either 0.18%dodecylmaltoside (“Formulation A”) or no dodecylmaltoside excipient(“Formulation B”) and a final sumatriptan concentration of 20 mg per 100microliter spray. Final pH adjustment is made using sulfuric acid orsodium hydroxide solution. A third formulation, Imitrex® sumatriptannasal spray, 20 mg per 100 microliters, manufactured by GlaxoSmithKline,is designated “Reference”. Each drug solution is administered to 18patients in a three-way crossover study, with a washout period betweendoses of at least 3 days, as a 100 microliter metered nasal spray usingstandard metered nasal spray devices such as those manufactured by Ing.Erich Pfeiffer GmbH, Radolfzell, Germany, Valois Pharma, Le Neubourg,France, or Becton Dickinson, New Jersey, USA. Blood samples arecollected from each patient at the timed intervals, for example 0.08,0.17, 0.25, 0.33, 0.42, 0.6, 0.67, 0.83, 1, 2, 3, 4, 6, 8, 12, 24 hours,as shown in FIGS. 14 and 15, for preparation of plasma from each bloodsample using K₂EDTA (dipotassium EDTA) as the anticoagulant. Plasmalevels of sumatriptan are determined by high performance liquidchromatography (Ge, Tessier et al. 2004).

FIG. 14 shows a comparison of the average plasma levels of all patientsat the various time points as indicated, along with the standarddeviation, for the Imitrex® nasal spray reference and Formulation Bwhich contains no alkylsaccharide excipient. The reference and theformulation B yield approximately equal performance with a Cmax ofapproximately 15 ng per mL and a Tmax of 1-2 hours.

FIG. 15 shows a comparison of the average plasma levels of all patientsat the various time points as indicated, along with the standarddeviation, for the Imitrex® nasal spray Reference and Formulation Awhich contains 0.18% dodecyl maltoside excipient. The Cmax forFormulation A is approximately 60 ng per mL, or approximately 4 timesthe Cmax observed for the Imitrex nasal spray Reference. The T-max isreduced from 1-2 hours for the Reference to approximately 8-10 minutesfor Formulation A which contains alkylsaccharide and the presumedtherapeutically relevant Cmax value of 15 ng/mL achieved by theReference formulation in 1-2 hours was reached in approximately 2minutes in the case of Formulation A.

The following parameters are calculated from the data obtained:

-   -   AUC_(0-t)=the area under the drug plasma concentration versus        time curve, from time of administration to the time of last        measurable concentration.    -   AUC_(0-∞)=area under the drug plasma concentration versus time        curve, from time of administration to infinity.    -   C_(max)=maximum drug plasma concentration.    -   T_(max)=time to the maximum drug plasma concentration.    -   Half-life=time after administration until the drug plasma        concentration is half of its maximum concentration.    -   K_(el)=elimination rate constant.    -   The mean pharmacokinetic results are tabulated below:

TABLE XXI AUC_(0-t) AUC_(0-∞) C_(max) T_(max) Half-life K_(el) Sample(ng · hour/mL) (ng · hour/mL) (ng/mL) (hours) (hours) (hour⁻¹)Formulation A 111.95 114.7 60.51 0.17 5.24 0.18 Formulation B 75.5577.36 15.67 2 4.68 0.16 Reference 79.52 81.06 17 1.25 3.89 0.19 (Form. A÷ R) × 128.54 132.65 305.23 — — — 100 (%)

Mean Cmax data at individual time points such as 0.08, 0.17, 0.25, 0.33,0.42, 0.6, 0.67, 0.83, 1, 2, 3, 4, 6, 8, 12, 24 hours for all thesubjects are plotted in FIG. 15.

Example 21 Administration of Alkylglycosides with Opioids IncreasesBioavailability

An opioid is dissolved in 20 mM sodium acetate buffer, pH 5.5 containing0%, 0.02%, 0.05%, 0.1%, 0.2%, or 1.0% alkylsaccharide. Opioids generallyinclude compounds having the general structure of Formula 1 as definedherein, such as naltrexone, methylnaltrexone, naloxone, morphine,codeine, thebaine, oripavine, heroin, oxycodone, hydrocodone,buprenorphine and nalbuphine. Each set of drug solutions is administeredto six groups of eight rats each by 20 uL instillation to a single nareof each animal. 200 μl blood samples are drawn by orbital bleed over athree hour time period at 0, 5, 10, 15, 30, 60, 120 and 180 minutes.After the last blood sample is collected each animal is euthanized withCO₂. After blood collection, plasma is immediately prepared from eachblood sample using lithium/heparin as the anticoagulant. Plasma samplesare stored at −70° C. until analyzed. Plasma drug levels are determinedby HPLC using the method described by Boulton or similar HPLC method.The concentration vs time data are plotted to determine the Cmax at eachalkylsaccharide concentration and the ratio of Cmax with and withoutalkylsaccharide present is calculated and will be recorded in the formatshown in Table XIX of Example 19. The observed Tmax values from eachplot will be determined by inspection of the concentration vs time plotsand will be recorded as shown in Table XX of Example 19. Opioids areexpected to exhibit the same or similar pharmacokinetic andpharmacodynamic parameters as triptans as described in Example 19.

Example 22 Nasal Administration of Opiods with Alkylglycoside IncreasesBioavailability and C_(Max) and Speeds Onset of Systemic Absorption

An opioid is dissolved in phosphate buffer containing eitherdodecylmaltoside (“Formulation A”) or no dodecylmaltoside (“FormulationB”) as described in Example 20, with a final opioid concentration of 20mg per 100 microliter spray. Opioids generally include compounds havingthe general structure of Formula 1 as defined herein, includingnaltrexone, methylnaltrexone, naloxone, morphine, codeine, thebaine,oripavine, heroin, oxycodone, hydrocodone, buprenorphine and nalbuphine.Final pH adjustment may be made using sulfuric acid or sodium hydroxidesolution. Each drug solution is administered to 18 patients in acrossover study, with a washout period between doses of at least 3 days,as a 100 microliter metered nasal spray using standard metered nasalspray devices such as those manufactured by Ing. Erich Pfeiffer GmbH,Radolfzell, Germany, Valois Pharma, Le Neubourg, France, or BectonDickinson, New Jersey, USA. Blood samples will be collected from eachpatient at the timed intervals, for example 0.08, 0.17, 0.25, 0.33,0.42, 0.6, 0.67, 0.83, 1, 2, 3, 4, 6, 8, 12, 24 hours for preparation ofplasma from each blood sample using K₂EDTA (dipotassium EDTA) as theanticoagulant. Plasma levels of opioid are then to be determined by highperformance liquid chromatography (Ge, Tessier et al. 2004).

Opioids are expected to exhibit the same or similar pharmacokinetic andpharmacodynamic parameters as triptans as described in Example 20 and asshown in FIGS. 14 and 15 as well as Table XXI.

Throughout this application, various publications are referenced. Thedisclosures of these publication in their entireties are herebyincorporated by reference into this application in order to more fullydescribe the state of the art to which the invention pertains.

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Although the present invention has been described with reference tospecific details of certain embodiments thereof in the above examples,it will be understood that modifications and variations are encompassedwithin the spirit and scope of the invention. Accordingly, the inventionis limited only by the following claims.

What is claimed is:
 1. A method of treating a condition ameliorated byepinephrine in a subject, said method comprising intranasallyadministering to the subject in need thereof a composition consistingof: a) epinephrine, b) an alkylglycoside, and c) inactive excipients,wherein the alkylglycoside has an alkyl chain including between 8 to 20carbons and the alkylglycoside concentration is between about 0.001% and10.0% (w/v), wherein the composition is an aqueous solution formulatedfor intranasal delivery to the subject.
 2. The method of claim 1,wherein the alkylglycoside is selected from the group consisting ofundecyl maltoside, dodecyl maltoside, tridecyl maltoside, tetradecylmaltoside, sucrose mono-dodecanoate, sucrose mono-tridecanoate, andsucrose mono-tetradecanoate.
 3. The method of claim 2, wherein thealkylglycoside is dodecyl-beta-D-maltoside.
 4. The method of claim 1,wherein the alkylglycoside concentration is between about 0.05% and 0.5%(w/v).
 5. The method of claim 1, wherein the composition provides a Cmaxfor the epinephrine in the subject that is about 2 fold or greater ascompared to administration without alkylglycoside.
 6. The method ofclaim 1, wherein the composition provides a Tmax for the epinephrine inthe subject that is about 2 fold or less as compared to administrationwithout alkylglycoside.
 7. The method of claim 1, wherein thecomposition provides a Tmax for the epinephrine of about 0.3 hours orless in the subject.
 8. The method of claim 1, wherein the compositionhas a pH of about 2.0 to 5.0.
 9. The method of claim 1, wherein theinactive excipients are selected from citric acid, sodium citrate,propylene glycol, glycerin, ascorbic acid, sodium metabisulfite,ethylenediaminetetraacetic acid (EDTA) disodium, benzalkonium chloride,sodium hydroxide, or a combination thereof.
 10. The method of claim 1,wherein the inactive excipients are selected from benzalkonium chloride,EDTA, or a combination thereof.